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We narrowed to 1,010 results for: PLE

Showing: 401 - 420 of 1010 results

  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. 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...

  4. 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...

  5. 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...

  6. 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...

  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. 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...

  9. 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...

  10. 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...

  11. 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...

  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. 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...

  14. 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 ...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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....
Showing: 401 - 420 of 1010 results