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We narrowed to 1,010 results for: PLE

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  1. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...high and above what would be expected in patient samples (e.g.: blood, urine). Adapting toehold switches...of the emerging outbreak, as many as 2.2 billion people globally live in areas conducive to the spread ...involved definitely contributed to our ability to complete the project on such a short timescale. Our article...pathogen’s genome as unique barcodes to identify samples containing the virus. Of the 48 sensors, targeting... diagnostic results.  Our approach now detects samples at clinically relevant concentrations (1-3 fM; ... look at viral load in patient blood and urine samples; we showed that we can detect the virus directly...Interview with Previous Collin's Lab Member and Sample 6 Cofounder Michael Koeris Learn How Minigenomes...

  2. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...structure of the hairpin-DNA complex is thought to destabilize the transcription complex, initiating cleavage ...Polyadenylation is the post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger...polymerase terminates differently. Polymerase III, for example, relies on a specific sequence and RNA secondary...mechanism, whereas Polymerase II termination is more complex and involves two RNA polymerase-associated proteins...polyadenylation enzymes, in a process that seems to couple termination with polyadenlyation. Mammalian expression...PMC104046. Wodrich H, Schambach A, Kräusslich H-G. Multiple copies of the Mason–Pfizer monkey virus constitutive...

  3. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...never enjoyed grant-writing. The more I talked to people with industry experience, the more I heard that...decision, the transition felt scary. While I knew two people from graduate school who were at AstraZeneca and...industry for any length of time, you’ll know more people in industry, which means you’re more likely to ...decade of work. The number of steps, decisions and people involved is staggering. The process is complicated...individual could not bring a drug to the market, so people in industry have to work together. Even as a postdoc...postdoc, you’ll have to work with other people in your group, and they are unlikely to hire you if they... academia, promotions in industry are based on people’s ability to manage others and work as a team as...

  4. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR. Human... helper viruses like adenovirus (Ad) or herpes simplex virus to facilitate efficient replication. The ..., and the two helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1:...quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet digital PCR (ddPCR) (Lock et al., 2014), or determination...known titer on every qPCR run. Note that the rAAV sample has to be digested with DNase before undergoing... Multiple Authors...

  5. Pooled Library Amplifications

    Type
    Blog Post
    ...library from Addgene, they’ll need to amplify the sample for further downstream applications. As individual...make the library more difficult to work with. For example, a library in a lentiviral backbone might be more... as individual plasmids in your library. As an example, if you want 100x coverage of your library, and...transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate transformation...libraries that are similar in size and composition as examples. Table 1 provides an overview of pooled library...protocols you can use as a reference. Table 1: Examples of Different Pooled Libraries at Addgene.  ...Libraries (PDF, 401 KB) Screening Zhang Lab - Multiplexed Overexpression of Regulatory Factors (MORF) Library...

  6. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...endogenously expressed SCF complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added...fusion protein with the endogenous CRBN E3 ligase complex which will polyubiquitinate the fusion protein....target protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation...acid (IAA), the TIR1 F-box associated E3 ligase complex will ubiquitinate the AID-tagged protein which ... Chung HK, Jacobs CL, Huo Y, Yang J, Krumm SA, Plemper RK, Tsien RY, Lin MZ. Tunable and reversible drug...Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc...

  7. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...millions of new cancer cases arise and millions of people die from cancer-related complications each year...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...cancer research has focused on picking apart the complex cellular pathways involved in the development of...pathway. Its major appeal is that it enables the simple construction of vectors to study the function of...transcript most commonly found across all TCGA tumor samples is represented, making the results of any such ...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...continue to to build and develop this new resource! Please leave us your thoughts in the comments section ...

  8. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...REPAIR and RESCUE can function in a multiplexed manner to edit multiple RNAs. It’s clear that CRISPR RNA ...research. RNA editing advantages RNA editing has multiple advantages over more traditional DNA editing systems...This change specifies the edit to be made when multiple As are present in the gRNA spacer, as ADAR will...target window, possibly due to longer stretches of duplexed RNA. Using an ADAR2DD catalytic mutant, they showed...By mutating residues in ADAR2 that interact with duplex RNA, they hoped to destabilize ADAR-RNA binding...As discussed above, the ability to edit RNA has multiple advantages, including reversibility and use in...

  9. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...years ago. Teaching by example No examination of the state of science would be complete without the inclusion...attempts to answer it in five paragraphs, with examples. If that sounds ambitious, that’s because it is...successful scientists have outgoing personalities to supplement their intellectual prowess and scientific skills...Four, “New Science,” Zimmer tackles more recent examples of scientific advances, and posits the way science... much less familiar with as a biologist--as an example of “big science,” a massive, expensive project ...certainly chapters that could warrant their own books. Multiple books have been written about CRISPR alone, and...

  10. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...targeting a specific genomic locus, which can require multiple cloning steps (Fujita et al., 2013). CRISPR-mediated...engineered DNA-binding molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii...another "in vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and ...interest with dCas9 and a targeting gRNA. One such example by the Xu lab is CRISPR affinity purification in...purification of a specific genomic region is another example of the many tools based on targeting of a genomic...genomic locus through a specific gRNA-dCas9 complex. When combined with next-generation sequencing, this ...research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii. References...

  11. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...mediate cleavage rather than the multi-subunit complex employed by class 1 systems. A putative new class...Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence requirements...than following it. The Cpf1 crRNA is also much simpler in structure than Cas9’s; only a short stem-loop...CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest to adapt to genome...vectors. In 2017, Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express...varies widely with gRNA sequence. Both groups used multiple methods to examine Cpf1 off-target editing. First...publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex Gene Editing by CRISPR–Cpf1 Using a Single crRNA...

  12. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    ...good guidelines to follow in order to ensure that people can quickly and easily identify what your plasmid... empty backbone name in your plasmid name. This simple piece of information can often convey many important...insert abbreviation to specify what species it is. Example: ‘h’ is for Human (homo sapiens), ‘m’ is for mouse...plasmid and their relative position to the insert. Example, if you have a Flag tag on the N-terminal of your...directly before or directly after the insert name.   Example: If the Glutamine at position 295 was mutated to...pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP These simple rules allow any scientist to know what is in a ...

  13. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    .... The optimized plasmid curing method was much simpler and took only ~32 hours instead of ~60 hours. ...strategy for plasmid curing, providing users with a complete editing process. The MUCICAT system While the ...including concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the ...Ptet, and Para. The pDonor plasmid pRE57-Ter with multiple terminators can serve to interrupt genes (or gene... pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction...dosages to facilitate swift optimization of strains. Multiple induction systems to choose from. Ready for metabolic...mainly on the development and application of a multiplex genome editing system based on CRISPR-associated...

  14. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...whole-genome/transcriptome” studies, in which, for example, the activity of all genes in response to an experimental...the globe (Nüsslein-Volhard et al., 2012). One example (which I will come back to) is the one-eyed pinhead...morpholinos. When injected into zygotes, morpholinos complementary to a target mRNA inhibit its translation either...genome is useful for a variety of applications. For example, specific tissues such as blood cells or neurons...regulatory systems have also been successfully implemented in zebrafish. Developmental biology insights ...study profiled 38,731 cells from 694 embryos over multiple stages and reconstructed the patterns of gene ...well-suited for neuroscience investigations. For example, neuronal activity can be tracked in whole brains...

  15. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...surface) of the leaf. This is a simple and easy technique to implement. Success might vary between experimenters...tobacco, Agrobacterium infiltration into leaves is a simple and efficient way to overexpress a gene and evaluate...your experiment (de Ruijter et al., 2003). For example, we did our initial transformations with the GUS... on transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase) with...temporal reporters to measure promoter activity, for example. Useful positive controls could include: pSB96 ...expression from Agrobacterium, but not the plant. For example, we have seen strong expression of the GUS gene...

  16. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...chalk talk? On the surface, a chalk talk is quite simple. It's you, a dry erase board, some markers (chalk...question, not a vague goal or a "what" question. For example, "we want a better understanding of the Z pathway...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...scientists outside your field, and interesting. For example, “there is a significant need for healthy livers...explain why it’s significant and innovative. For example, "while it is possible to grow liver stem cells...chalk talk Perfect your story and delivery. Design simple figures and diagrams to illustrate your vision....also helpful to practice at least a few times with people that are outside your lab and outside your field...

  17. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...designated this fragment as the ccdB region (for “coupled cell division”). The encoded toxin-antitoxin system...subunit of DNA gyrase and trapping it in the cleaved complex. This results in DNA breakage and cell death. Conversely... so that the marker can be reused to introduce multiple genetic modifications in the same strain. MazF...translation. Here, Mok and Hok translation is coupled. Translation from the Shine-Dalgarno (SD) sequence...often lost without plasmid addiction systems. For example, Cara Morin and James Kaper used the hok/sok system...Teru, and Sota Hiraga. "Mini-F plasmid genes that couple host cell division to plasmid proliferation." Proceedings...K. Burley. "Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition....

  18. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...blood sample, in the best-case scenario [7]. In the clinical setting where patient blood samples are limited...especially if used to make clinical grade hiPSCs. For example, the CytoTune™-iPS 2.1 Sendai Reprogramming Kit...persistence of residual viral material [6]. The complete removal of the residual viral particles and establishment... in the presence of these two factors are not completely understood, the addition of an shRNA for P53 ...grade hiPSCs from easily accessible patient blood samples. All pCXLE vectors listed below in Table 1 are ...cells from CD34+ cells across blood drawn from multiple donors with non-integrating episomal vectors. ...

  19. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...of CRBNmidi alone (A) and in a ternary complex with the multiple myeloma drug mezigdomide and ubiquitination...Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using...on its own, with drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as ...mutations.  Here, the researchers were able to target multiple genes in M. abscessus either simultaneously or... (also cyan) embedded in the Titin I10 protein (purple) only when Titin I10 unfolds under tension. Upon... Multiple Authors...

  20. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...maintain itself within host cells.  The plasmid has multiple ori that are functionally differentiated from ...proteins from bacterial hosts. In a plasmid with multiple oris, one may function in one type of host and... the others are functional in other hosts. For example, the pJD4 plasmid harbors distinct origins (ori1... load (Meyer 2009). Figure 1: Idealized examples of a narrow host range plasmid with only one ori... ori and a broad host range plasmid with multiple oris that vary in DnaA-box and iteron structure, host-independent...Acinetobacter calcoaceticus, Actinobacillus pleuropneumoniae, Actinomyces naeslundii, A. viscosus, Aerobacter... hosts in which proteins can be expressed. For example, scientists can incorporate interchangeable antibiotic...

  21. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ... relevant bases within the editing window, for example all C’s will edit to T’s. Thus, it’s important ...knocking out a particular gene isn’t feasible. For example, temporary repression or knockdown allow you to... target genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused...SunTag system which facilitates recruitment of multiple activation domains to the same genetic locus via... proteins. The Synergistic Activation Mediator complex, generally the strongest activator, consists of...Edit”. Is Cas9 not your Cas of choice?  There are plenty of other enzymes to choose from within the Cas ... Multiple Authors...

  22. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...the progress of the project. After that rather unpleasant experience I thought: I could do this job so ...companies produce different products and therefore need people with different areas of expertise in technical .... In addition, it can be advantageous to speak multiple languages - that makes communication with customers...are different names for technical support roles. Examples are Technical Service Representative, Technical...optimize” a particular standard lab protocol. For example, someone may attempt super quick incubation times...product in question. This can solve the problem in a simple way even if you don’t exactly know why. Personal... use these communication skills by working with people from many different backgrounds and realizing that...

  23. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ...consultants who enjoy their jobs most, get pleasure from taking up such complex challenges, quickly breaking them...got my PhD a couple of years ago in Bioengineering, and one of the things that pleasantly surprised me...Healthcare consulting is a fast-paced field, requiring people who are willing to quickly learn, and apply their...The types of projects mentioned above are just examples, and healthcare consulting firms can be involved... career in several different fields. While some people continue to enjoy the challenges of healthcare ...

  24. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...coming weeks, but, in this post, we will outline a simple method for selecting the guide RNA, validating ...CRISPR Design tool supplied by the Zhang lab. This simple website invites you to input the target region ...happy with your selection, ordering the sgRNA is simple and easy. Several companies supply gRNA sequences...you gRNA in vitro first. In vitro screening is a simple and fast method for validating your selected gRNA...). Testing the efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your ...perfect primer sets to use to genotype your pups. A simple PCR with these primers will let you know if there...-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering...

  25. The Challenges of Cell Culture

    Type
    Blog Post
    ...put, the majority of cell line problems come from people sharing cells: the black market of cell biology...when it is important to check their identity. For example, perhaps a working cell bank has been made by a...stocks of cells. Working stocks should be frozen in multiple vials and thawed to produce a new batch once you...the cells you are working with because you have plenty of maintained freezer stocks. It is vital that ...monitor any changes. Your tracking system could be as simple as a piece of paper on the front of the incubator...passage number, cell line, and date, which you complete each time you passage the cells (as well as recording...are changing but yet more is needed to convince people of some of the basic tenets of responsible cell...

  26. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ... for using lentivirus, infect and select, seems simple, in actuality, many find using lentivirus to be...al. outline a do it yourself PCR-based approach complete with freely available positive controls. Routinely...labs prefer to collect and pool the harvests from multiple time points while others opt for a single harvest...transfer and packaging plasmids. A benefit of the multiple harvest strategy is that each day you are supplying...destabilize the viral envelope. One drawback of the multiple harvest approach is that it tends to result in...buffers used, and other experimental parameters. For example, while freeze thaw is generally not recommended...that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, it is best to...

  27. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...proteins. lon protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce...eukaryotic proteins Lemo21 (DE3)* Chloramphenicol (pLemo) Rhamnose-tunable T7 RNAP expression alleviates...replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo (CamR) Expression of toxic, insoluble, or membrane...variety of sources, so fear not—there are a few simple troubleshooting measures that can help get you ...will not express in an IPTG induction strain for example, nor will a p15 plasmid be compatible with a pLys...cell-density they support; however, minimal media supplemented with M9 salts may be preferable if the protein... Proteins which contain hydrophobic regions or multiple disulfide bonds may aggregate and become insoluble...

  28. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    ...a world of possibilities in the engineering of complex gene networks. A comparable system for targeting... of RNA is highly desirable for extending the complexity of genetic circuits, allowing for tighter spatio-temporal... of interest. The new tool was developed and implemented by applying the Golden Gate cloning method to...March is fused to a translational repressor. In principle, however, designer PUF proteins can be fused to...in addition to their use in basic biology, for example, by targeting pathways underlying cancer or other...Assembly Kit out for themselves, she does have a couple of tips: First, she recommends using standard BsaI...

  29. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...Institute for Biological Studies) Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum...spectrum is perfect for use in spectral multiplexing (Shcherbakova et al., 2018). NIR FPs can be combined ...Verkhusha, V.V. Near-Infrared Fluorescent Proteins: Multiplexing and Optogenetics across Scales. Trends Biotechnol...11, 239 (2020). Oliinyk, O.S., Shemetov, A.A., Pletnev, S., Shcherbakova, D.M. & Verkhusha, V.V. Smallest...cyanobacteriochrome as versatile tag for spectral multiplexing. Nat Commun 10, 279 (2019). Oliinyk, O.S. et...Huisman, T.M., Verkhusha, V.V. & Hodgson, L. Direct multiplex imaging and optogenetics of Rho GTPases enabled...

  30. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...locations using Cas9 and track the localization of the complex. Broccoli, the other green fluorescent aptamer,... RNA. Fluorescence is observed when these RNAs complex and the entire aptamer can hybridize and fold. ...uniqueness of the photostability of the Corn-DFHO complex. For dessert (last but not least), the only fruit-named...fruit-named aptamer, Mango, binds to multiple derivatives of thiazole orange (TO) and fluoresces in the...Other groups have used Mango to study RNA-protein complexes (Panchapakesan et al., 2017) and short non-coding...mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology...transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017): 1187. PubMed...

  31. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ...resources on self-promotion (how hard it is for some people, especially women), visibility (how to get it, ...especially if introverted ), and networking (how to get people to remember you). What better way to accomplish...Interviews: The All-Important Job Talk. 1. Watch other people give great talks We are not talking about lab ...simplicity of their slides and how the slides complement the spoken message. Learn from their level of...cause and effect. Stories can and should be very simple. The goal is to get the listeners to be thinking...on a slide but not reading the explanation. The people that need the information will find it. Experienced...particular question. I like to ask about how many people use plasmids in their research and then see how...

  32. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...Brainbow multicolor labeling system - a perfect example of the continued utility of Cre-lox. Check out ...events specified by three loxP variants, but the principles remain the same. The first fluorescent protein...than 3-4 colors due to the tandem integration of multiple copies of the construct (about 8 in Brainbow-1.0...-1.0 mice.) The combinatorial effect of multiple, independent recombination events leads to a rainbow ...Brainbow-2 each contained a “default” state; for example, Brainbow-1.0 expresses RFP when the construct ...system, Sanes and Lichtman also developed a complementary Flpbow construct that is functionally similar...13):2835-46. doi: 10.1242/dev.094631. PubMed. Multiplex cell and lineage tracking with combinatorial labels...

  33. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ... by multiple keywords that are separated by a single space to get even more specific. For example, to ...e.g. SNPs) exist. Additionally, consider testing multiple gRNAs targeted to the same locus to ensure their... are not generated from a plasmid. If you, for example, prefer to deliver your CRISPR components as RNPs...and you can search by any part of the data. For example, entering ‘C. elegans’, or ‘elegans’ will filter...publication. As you develop and confirm new gRNAs, please consider submitting their sequences (and plasmids...

  34. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...DePace’s yearly planning meetings revolve around two simple worksheets, each of which is filled out separately...summarized briefly below (and available of in the supplement of this paper), help each person organize his...including major project goals and effort needed to complete them. These documents dictate the subject matter...concrete plan in hand, trainees are motivated to complete their work rather than overwhelmed or confused...meetings may not look the same in every lab, but the principles behind this technique are beneficial for researchers...Extracurricular Activities for a Strong Science Career Path 7 Simple Strategies to Resolve Conflict With Difficult Supervisors...

  35. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ... and a mashed potato bar (complete with all the fixings), as well as plenty of delicious desserts. We ... 10-year Anniversary Gala celebrated all of the people who have helped Addgene grow and spread our mission...playing all night was hand-picked (like our plasmid samples!) by Addgenies. If you were there, you might have...Addgene over the last 10 years as well as the many people that make up the Addgene family. It was great to... it to fruition. The concept, though relatively simple, changed the course of scientific research. What...

  36. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...sensors Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV vectors are common...vectors, and post-translational regulatory elements. Examples of these approaches are given in Tables 3-7. 3...specificity of cell targeting even further. For example, injecting AAVs at one location can result in expression...that received information from a region. In the example shown in Figure 1, injected retrograde AAVs at ...neuroscience is becoming more and more versatile. To complement the wealth of genetically encoded tools available...only reagents, but also method information. To complement these efforts, we’re beginning to share data ...

  37. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...Scientists are excited by somewhat unusual things. For example, I am not that interested in meeting movie stars...astronaut Jim Lovell I was speechless (the club of people who have been in space is pretty small). Therefore...this over 30 minutes, which was fun to watch. A simpler 1 protein - 1 RNA system Drs. Dounda and Charpentier...into one RNA to direct cleavage the result was a simpler 1 protein - 1 RNA system.  Dr. Charpentier advocated...breakthrough. In fact, the panelists agreed that, for example, the technology is already available today to cure... anemia in stem cells but this can’t be used in people until we learn more about safety and more about...

  38. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...engineered ESCs), and chimeric adults mated to produce completely transgenic offspring. Although powerful, this...about 100 bps into the run, it got “dirty”, with multiple sequence traces overlapping each other! When the... finally, we have a technology, CRISPR, that is simple in concept, straightforward to use, and robust ...missing or added at the broken ends and, as such, cripple the gene. However, if repair material is provided...enrichment and selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating... Wu X, Jiang W, Marraffini LA, Zhang F (2013) Multiplex Genome Engineering Using CRISPR/Cas Systems. Science...-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering...

  39. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    ... or condition of interest using a simple keyword search. For example, searching the term "breast cancer...results section, but you had to dig through the supplemental files to see the actual data. Now you’re wondering...datasets, which are often obscured in large supplementary data files, BioGRID ORCS makes it possible for...sgRNA-level sequence read scores. The original supplemental data tables available with the publication are...broadest possible scope of screens to be curated, for example phenotype screens with read-outs other than cell...

  40. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    Blog Post
    ...built, literally, 'brick-by-brick' to introduce people all around the world to the lives of researchers...wrong. Whether it be a bad fridge freezing your samples, or a dysregulated water bath boiling your experiments...Unfortunately, unless they’ve worked in a lab, most people don’t get to experience this very human side to... is to help make science mainstream and to show people the very human struggles in the laboratory routine...sharing and laughing at these struggles will bring people together and make these daily challenges a little...that we step on them with bare feet), to connect people with science. And so, “Bricking Science” was born...

  41. An Addgene Summer Internship

    Type
    Blog Post
    ... had to get used to the workspaces, as well as complete the lab safety and cybersecurity training before...project is a major project given to each intern to complete and present on.    Figure 1: Addgene's...weeks to work on cloning the Syntaxin3 gene in a pLenti vector backbone. During those five weeks, I worked...obtain eight clones. We ran digested and undigested samples of the clones on an electrophoresis gel in order...a 100% successful clone of the Syntaxin3 in the pLenti backbone! Once everything was confirmed, I worked...Toj-Villagran is a rising senior in high school who just completed their summer internship at Addgene. They plan ...

  42. The Blue Flame Award: Celebrating Addgene's Most Requested Depositors

    Type
    Blog Post
    ...can gauge if people are happy depositors is if they deposit plasmids more than once. People get hooked ...Flame Awards. What's in a flame?     We’re always pleased when scientists around the world choose to share...Sainsbury Laboratory in Norwich, UK: “We’re very pleased to distribute our plasmids via Addgene. Given the... plasmids and have not yet received your award, please contact [email protected] to claim your Blue Flame...deposit. Begin yours here. It has been my genuine pleasure to send these small tokens of thanks for the incredible...

  43. Celebrate Open Data Day with Addgene’s AAV Data Hub!

    Type
    Blog Post
    ... detection methods. For example, you can provide location, titer dose (Example: 1E13 GC/mL) and subject...this resource for the community. There are a few simple steps you can take to input your data for sharing...and assessment of your experimental results. For example, if you visualized expression after AAV injection...generated with a caption. For microscopy images, please include channel imaged, a color legend for probes...clear overview of what you need to submit data. Please note that the data hub is currently still in beta...

  44. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ...until you don't, of trying to understand your completely unexpected findings, or understanding why your...thrice-failed experiment, this paper was actually from a completely unexpected finding in a different experiment,...your conflict, you’ll need two other things to complete your story: an introduction, and a resolution.... big-picture problem, by asking your audience a simple question, like “Have you ever thought about how...conflict to get your audience invested in the more complex story, and then build on that in your resolution...journey, stories from patients or other affected people, man/animal vs nature, anthropomorphized animals...

  45. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    ...educational materials to our repository of open samples. We are, and have always been, a community resource... to maximize the reach and impact of your work. Please email [email protected] if you would like to ...ideas for new materials to add to the collection, please let us know by filling out this suggestion form... will find useful antibodies for your research. Please consider sharing your experiences with antibodies...recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain. eLife 8:e43322. https....43322 Bandrowski, AE, Martone, ME (2016) RRIDs: A Simple Step toward Improving Reproducibility through Rigor...

  46. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ..."CellTag Indexing: genetic barcode-based sample multiplexing for single-cell genomics." bioRxiv (2019)... be tagged with different CellTag libraries at multiple timepoints during reprogramming. These library-specific...library-specific identifiers act like a timestamp and help demultiplex the scRNA-seq data and build multi-generational...phenotype typically emerges. Every three days, a sample of cells was collected for scRNA-seq analysis and...tended to have similar reprogramming outcomes. For example, cells with shared CellTagD13 tags often had similar...

  47. Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

    Type
    Blog Post
    ...say that most people do not recognize its importance or perhaps even know it exists. Couple that with a ...in Columbus, Ohio, ABRC ships more than 100,000 samples to researchers and educators in 60 countries each... resources, we seek out opportunities to engage people in science learning. Whether visiting classrooms... 2017 alone, ABRC staff reached more than 4,000 people through our outreach efforts. Our education partnerships...Addgene.org Find Plant Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants...

  48. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    ...would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am ... mentors -- well, it's time you call upon these people for guidance.  Before even beginning the process...this part might hurt, but if you pitch to enough people, you will find the answer to your viability question...asking to meet. I have cold emailed hundreds of people asking for meetings to discuss miLab.  Although...impressions are everything and you want to leave people remembering your potential as an entrepreneur, ...

  49. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ... in principle adapting a FRET sensor for monitoring calcium versus zinc should require the simple swapping...dozens of other published FRET-based sensors. For example, his team is now applying it to improve upon an...domains, in practice optimizing the sensors involves plenty of trial and error. Transition metals also present... of its kind. “I hope by depositing at Addgene, people will start using them,” Merkx said. “For us, it... developed something useful [enough] that other people start using it. For us, it’s like a test.” References...

  50. Interview: Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering

    Type
    Blog Post
    ...and what it can enable? Boyden: There are many examples where you want to activate two sets of neurons...humanistic and philosophical implications. As an example, a group at Caltech used an optical fiber together... to treat patients? Over a quarter of a million people have some kind of neural implant already. If we...Those are two of the biggest ones. And as a third example, it would be helpful to have molecules with better...three or four hour conversation. The brain is very complex and we've hosted hundreds of visitors in our lab... different from a cell in a dish. I always tell people, 'Be driven by your science.' Looking for Optogenetics...
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