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We narrowed to 1,018 results for: REN

Showing: 481 - 520 of 1018 results

  1. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...Insertion of the mini-transposon can occur in two different orientations, but is strongly biased towards T-RL...integration efficiencies achieved with VchINT, at two different incubation temperatures. Data adapted from Vo ...and R ends - thus, scarless insertions are not currently possible, unlike methods involving homologous ...thanks to our guest blogger Leo Vo! Leo Vo is currently a PhD candidate in the Sternberg Lab at Columbia...Columbia University Medical Center. He currently studies CRISPR biology and CRISPR genome engineering technologies...Message or follow him on twitter @Vopinator.     References: Halpin-Healy TS, Klompe SE, Sternberg SH, Fernández...

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

    Type
    Blog Post
    ..., and he wisely makes no such attempt. He is transparent about his own biases--many of which will be shared...journal editorials, science social media, and conference workshops for years. That is, presumably, why... is the title of the chapter that examines the inherent brokenness of scientific publishing and research...off the section saying “science itself is never inherently bad or evil. However, it is easy to do sloppy...today--the day you are reading this review--in the current period of rapid flux, the chapter might still feel...scientist who enjoys learning a little bit about many different things (part of the reason Addgene has been such...

  3. An Introduction to Adenovirus

    Type
    Blog Post
    ...written by Addgenie Brian O'Neill.  References and resources References Bauzon, M., Castro, D., Karr, M.,...three (Servellita et al., 2023). Normally when children have acute viral hepatitis it is caused by the... the removal of certain adenoviral genes that apparently contribute to virulence (Tollefson et al., 1996...B. (2014). Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https...Y. (2023). Adeno-associated virus type 2 in US children with acute severe hepatitis. Nature, 617(7961)...

  4. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...bottleneck in the experimental process. The majority of current cloning techniques involve the assembly of a circular...single PCR, for example for subcloning genes from different templates. All PCRs are run as an 18 cycle, 25... - 1 μl DpnI is added to the PCR mix to remove parental DNA (37 °C for 15 mins). This enzyme specifically...the MRC Laboratory of Molecular Biology. He is currently investigating the molecular biology of synaptic... on Twitter @jakefwatson and @JGarciaNafria.  References 1. Jones, Douglas H., and Bruce H. Howard. "A...for plasmid cloning Find our molecular biology reference here Find molecular biology protocols here   ...

  5. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...of green fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly, cell biologists...dye (Figure 1a) and FAP systems (Figure 1e) are inherently fluorogenic and several others can be made fluorogenic... We continue to develop derivatives that have different spectral properties, exhibit high fluorogenicity...contributed by Luke D. Lavis. Luke D. Lavis is currently a Group Leader at the Janelia Research Campus,...modalities. Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. ...Imaging proteins inside cells with fluorescent tags." Trends in biotechnology 30.1 (2012): 8-16. PubMed PMID...

  6. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...moving around to explore the environment has a different motivation (and brain areas associated) compared...addiction. Anxiety tests Rodents have a natural preference to stay in dark areas (protected) as opposed ...associated with psychiatric diseases such as schizophrenia or bipolar disorder. In both diseases, animals...these in a  future blog post, so stay tuned!  References Alpár, Alán, et al. "Hypothalamic CNTF volume...volume transmission shapes cortical noradrenergic excitability upon acute stress." The EMBO journal 37.21 ....12 (2007): 1079. PubMed PMID: 17700574. Hare, Brendan D., et al. "Optogenetic stimulation of medial prefrontal...

  7. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ... lot of time. Don't re-invent the wheel. Test different leaf ages Usually younger leaves work better. ...luciferases to more easily quantify the effect of the different treatments on transformation success. Some reporter... all plasmids from the Lee-Parsons lab. Test different leaf infiltration methods Co-cultivation (Li ...in the plant but not from Agrobacterium. Test different Agrobacterium strains We used the disarmed Agrobacterium...allows you to quickly assemble vectors and test different ideas. We are a huge fan of the Golden Gate based...starting his work in the Lee-Parsons Lab.     References D'Aoust, Marc-André, et al. "Transient expression...

  8. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...Austrian physician, Andreas Rett, who tried to raise awareness and published his observations in a German medical...hope that a similar reversal may be possible for children and adults with Rett Syndrome.  Since these discoveries...discoveries, scientists have studied how different parts of MECP2 influence Rett Syndrome. Zoghbi’s lab...varying degrees determine the clinical severity of different MECP2 mutations. More recently, the Bird lab found..., Rett databases, and other helpful tools.  References: Amir RE, Van den Veyver IB, Wan M, Tran CQ, ...Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery. Molecular...

  9. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...using FACS which could limit their use across different cell types and systems. To introduce flexibility... GO works by affecting protein translation of different reporter proteins. Protein translation requires...used GO to induce the translation of an array of different reporters including mScarlet-I, luciferase, antibiotic...proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran.../10.1093/nar/gkaa124 Martin AS, Salamango DJ, Serebrenik AA, Shaban NM, Brown WL, Harris RS (2019) A panel...s41598-018-36739-9 St. Martin A, Salamango D, Serebrenik A, Shaban N, Brown WL, Donati F, Munagala U, ...

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

    Type
    Blog Post
    ...CRISPR-associated transposition.     References and Resources References: Jiang Y, Chen B, Duan C, Sun B, ... E. coli host, and the addition of a sacB gene renders it beneficial for plasmid clearance. It is more...concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the plasmids by...MUCICAT toolkit includes versions of plasmids with different induction systems, such as PT7, Prha, Ptet, and...Capability to construct a strain library with different gene dosages to facilitate swift optimization ...

  11. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi E,...development of tools that exploit and expand our current knowledge. The Abudayyeh-Gootenberg lab (also called... was sent to the AbuGoot lab for testing, they weren’t sure what they would find… but it was safe to say...fibrosis, which can be caused by thousands of different mutations, all found in the same gene. If we only...the number of potential applications. But they weren’t done engineering quite yet – surely, they thought...

  12. Analyzing CRISPR Editing Results with ICE from Synthego

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    Blog Post
    ...Synthego has developed a new tool called ICE (Inference of CRISPR Edits) to solve this problem. This tool...efficiency and determine the profiles of all the different types of edits that are present and their relative...compares hundreds of samples simultaneously.  The current version of ICE software can analyze indels that...Adjacent Motif (PAM) sequence for the nuclease used. Currently, ICE is configured for Cas9 fromStreptococcus ...average amount of signal that disagrees with the reference sequence derived from the control trace file. ...cut site, representing a high level of sequence difference. Traces The Traces tab shows the edited and ...

  13. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...example, CRISPR activation has been used to differentiate induced pluripotent stem cells (Chavez et al...neuronal cells by activating genes that cause differentiation and to reverse HIV viral latency (Bialek et...already be hitting an upper bound of activation that current Cas9 systems cannot help you pass. In addition,...directed towards that gene as there can be a large difference in guide potency. Thus, in cases where users ...Chavez et al., 2016). To learn more about the different types of Cas9 activators, check out our blog post...DNA sequence or regulating gene expression.     References Behan FM, Iorio F, Picco G, Gonçalves E, Beaver...

  14. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... to determine the pros and cons of using many different types of gRNA design software. When you are happy...yourself through whatever DNA supply company you currently use. You can also find many previously validated...inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a...weeks old and use the ear tag method (creating different patterns of ear holes and using that material ...latest and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target...plasmid into zygotes." Development, growth & differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541...

  15. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...tagged protein because of its size. This type of interference is known as steric hindrance. For instance, ...OSER whorls. This assay is quantitative and is currently the standard for assessing oligomerization in ...fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus. For...advisable to generate multiple constructs with different insertions, since it is difficult to predict which...can follow him on twitter: @joachimgoedhart.   References Adjobo-Hermans, Merel JW, et al. "Real-time visualization...291.5512 (2001): 2408-2411. PubMed PMID: 11264536. Klarenbeek, Jeffrey, et al. "Fourth-generation epac-based...

  16. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...similarities with humans. However, rats have some key differences in physiology, intelligence, and human correspondence... generate transgenic axolotls for an array of different studies. Elly Tanaka’s lab has constructed several...include understanding of allelic variation between different plant species and metabolic properties affecting... collections to help start your experiments! References: Blount, Z. D. (2015). The unexhausted potential...Zhang, C., & Xing, X.-H. (2018). Pooled CRISPR interference screening enables genome-scale functional genomics...Plasmids Collection Page Read our molecular biology reference Find plasmids for your research ...

  17. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different RTKs. Although...Improving on the glutamate sensor iGluSnFR from Loren Looger’s lab, Kaspar Podgorski and colleagues have...response. They generated variants optimized for different applications (v82 has higher affinity for glutamate...By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato...sub-millisecond resolution and sub-degree temperature differences under a variety of imaging conditions (155 μs...

  18. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...solubility, and thus different reagents are useful for biotinylation in different microenvironments. There...used today in molecular biology for an array of different techniques and protocols. In this post we will...determined to be due to a deficiency in Vitamin H later renamed Vitamin B7/biotin. Egg white injury would plague...a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular...post-translational modification of exceptional specificity.” Trends in Biochemical Sciences, 24.9 (1999b): 359–363....

  19. Replacing Paper: Tips for Choosing an Electronic lab Notebook

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    Blog Post
    ...labs. While many labs approach the situation differently, some adopting simple note-taking tools, others...functionalities with experiment records. Many different types of ELNs are available on the market today...available here. Our questions focused on researchers’ current practices, their opinions towards ELNs, and their...desired functionalities i.e. main priorities of different ELN features, ranging from whether respondents...cases this applies to universities. How are you currently exchanging information with your team? If you ... allows you to assign specific permissions to different lab members to avoid any issues later on. Do you...

  20. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ... iPSCs (or correct them back to wild type), differentiate them into somatic cells affected by the disease...iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones! We ... experiments with blocking mutations, we still weren’t entirely happy with our editing outcomes and in...Dominik Paquet and Dylan Kwart Dominik Paquet is currently a Professor for Neurobiology at the Institute ...Tessier-Lavigne’s lab at the Rockefeller University. He currently works to understand pathological mechanisms of...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering...

  21. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...scientists have developed and fine tuned many different ways to clone DNA fragments which have provided...with an enzyme master mix, which contains three different enzymes: an exonuclease, which chews back the... this doesn’t allow for mix and match of many different fragments. One way around this is to use a combination...with a G!) but actually work by significantly different methods. Golden Gate cloning does result in the...from one vector to another. There are lots of different ways to clone these days. Gibson and the other...biology, you should consider giving it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC,...

  22. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    ...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the monkey story hit the presses, another very different bit of CRISPR news was circulating. Scientists...how CRISPR/Cas9s actually work. Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales...this structure may help us engineer around the current limitations of the Cas9 complex," said study author...

  23. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...by Hannah Dotson in Dec. 2022.  References and Resources References Gossen M, Bujard H. Tight control...In the presence of tetracycline (Tet), TetR preferentially binds Tet over the TetO elements, allowing ... One can therefore place GAL4 and GAL80 under different promoters and create sophisticated patterns of...wondering, “why is it necessary to have all 3 of the different binary systems above if they all behave very similarly...tet-on-and-tet-off-2nd-generation Mullick, A., Xu, Y., Warren, R. et al. The cumate gene-switch: a system for...

  24. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin,...rearrangement (such as when a stem cell initiates differentiation), transgene silencing may occur (as it would...cassette can be incorporated, and used during the differentiation process. Moreover, a splinkerette-PCR assay...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-...piggyBac-ing models and new therapeutic strategies.,” Trends Biotechnol., vol. 33, no. 9, pp. 525–33, Sep. 2015...26299960. PubMed Central PMCID: PMC4559351. 8. A. G. Uren, H. Mikkers, J. Kool, L. van der Weyden, A. H. Lund...

  25. Science Communication for Everyone

    Type
    Blog Post
    ...Toronto, created The Addictive Brain to spread awareness about how scientists utilize tax-payer money and... how our articles have helped them understand different topics of science, especially disease related ...terms like manipulations, anomaly, and so on, differently than scientists would. For example, “manipulate...blogger, Chinmaya Sandangi! Dr. Chinmaya Sadangi is currently a postdoc at the University of Toronto and founder... receive our science career guide Learn about different careers in science communication Resources on...

  26. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...fragments that are very different.” Because the fragments produced will have different overhangs, they can...systems. “The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann...Lohmann of Heidelberg University. “What is different is the plasmids you use to bring this into your target...available, making it easier for us to do our work.” References 1. Athanasios Lampropoulos et al. "GreenGate ...

  27. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

    Type
    Blog Post
    ...5,000 different research institutions. This has given us a unique window into technology trends in the...by the influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids...HA-Ubiquitin-WT and HA-Ubiquitin, and we now have 19 different ubiquitin plasmids in the repository. 100,000 ... and modified by so many labs and for so many different purposes that we had to create a CRISPR page to...

  28. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    ...controlling properties like growth, survival, and differentiation.” https://www.addgene.org/kits/sabatini-wood-cancer-pathway-orfs...implicated in cancer cell proliferation, survival, differentiation, and apoptosis. For each of these 17 pathways...morphological features, migration, or survival in different contexts.” It’s also possible you could make use... sometimes drug screens and sometimes totally different experiments we never thought of - and that’s thrilling...constructs to the kit in the next two years. References 1. Martz, Colin A., et al. "Systematic identification...

  29. 15 Hot Plasmids from 2017

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    Blog Post
    ...tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community. Don E...PiggyBac transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered...is a new member of this class that works via a different mechanism - the PhoCl protein is irreversibly ...“caged” by fusion to specific domains thereby rendering them inactive in the cytoplasm. The authors use...interest. NHEJ- or HDR-induced mutations in ATP1A1 can render cells resistant to the small molecule ouabain -...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ...examining genomes possessing CRISPR systems with apparent self-targeting. Two of these Lmo proteins, AcrIIA2...

  30. Spurred by COVID-19, Addgene Bridges Plasmid Sharing Between Academia and Industry

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    Blog Post
    ... Conversations with children playing in the background became a normal occurrence as Addgene and our university...light of the need for basic research during the current pandemic, universities are showing an unprecedented...of Illinois Chicago                     Boston Children's Hospital                           California...scientists. This partnership complemented each other's strengths: Ginkgo Bioworks’ infrastructure allowed them ...

  31. Working 9-5: an Addgene Co-op

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    Blog Post
    ...year bioengineering student, to Addgene. I am currently working on the QC/QA team as a DNA Sequencing ... food service industry, which followed a very different schedule. Fortunately, I was provided with a detailed...gave me enough time to become familiar with the different types of issues that were important to this function...Specifically as a co-op, it is encouraged to explore different teams and work on projects there. At this point...point, I have seen several lab processes and am currently working on projects for the Science Communication...

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

    Type
    Blog Post
    ...neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists... populations using viral strategies Combining different approaches can increase the specificity of cell...optogenetic payload in neurons that project to different regions of the brain. A more locally defined set...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...share experimental design and data. With these different approaches to open science, we aim to accelerate...

  33. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three colors. The...specificity of these orthologs is key: due to differences in the PAM sequences required by each ortholog...specific for telomeric sequences and showed that different fluorescently labeled dCas9 isoforms are efficiently...target sequence. They were also able to label two different pairs of chromosomes using gRNAs specific to sequences...Addgene are excited to see what’s coming next! References Multicolor CRISPR labeling of chromosomal loci...

  34. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    ...trained to be good at learning new things—this is a strength you can draw on and demonstrate in your application...be excited to be there. If you are appying for diffrent types of positions (for example, bench research...resume should also be customized to highlight different skills and to get the important information in...hiring manager. Learn to confidently describe your strengths in your resume and don't be shy about asking for..., you will never get the help that can make a difference. Having your resume forwarded personally gives...

  35. An Addgene Co-op: The Intersection of Fun, Friendship, and Work

    Type
    Blog Post
    .... Northeastern compiles a list of hundreds of different jobs for students to apply to, and after a grueling...marketing and social media co-op. There are a lot of different worries that come along with entering the workforce...and co-founder, approached me and asked about my current projects and personal interests. We chatted about...participated in the discussions, I began to note trends in the data, and from that, suggest changes to ...of my co-op. I had to do research on loads of different best practices, create an email strategy, and ...

  36. Hot Plasmids - October 2022

    Type
    Blog Post
    ... tissue, in vivo. Two of their constructs are currently available as Cre-dependent, EF1ɑ-driven viral ...have high selectivity for K+ over Na+ and rapid photocurrent kinetics. They demonstrate the use of HcKCR1...two-photon excitation around 1,040 nm, has larger photocurrents, and is more selective for K+ than HcKCR2, making... be used as a potent optogenetic silencer. The current work also proposes promising avenues for future...anti-c-Myc [9E10] (isotype IgG2a) antibody is currently recommended for use in western blot, but we are...

  37. Antibodies 101: Affinity Tags

    Type
    Blog Post
    ...antibodies, and more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, ...Nonetheless, this tag can be purified with several different resin types and is a fairly small tag that is ...  Affinity tags vs. other tags There are many different types of tags, including epitope tags (FLAG, Myc..., D. W., & Wingfield, P. T. (Eds.). (2001). Current Protocols in Protein Science. John Wiley & Sons, Inc...

  38. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ... Substrate: Some luciferase reporters, such as Renilla luciferase (RLuc) or NanoLuc, require injection...together genes from gas vesicle operons from two different bacteria, Bourdeau et al created two constructs... 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles is the amount ... invisible to ultrasound, which allows for differentiation of arg1 and arg2 ultrasound signals. This also...and arg2 plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan...

  39. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    ...and insecticidal proteins to the development of renewable biotechnology products and biosensors for herbicides...Development of a bacterial expression system for renewable biotechnology products Dr. Idah Sithole-Niang,...database for this important agricultural plant. Her current research focuses on endophytes, bacteria, and fungi...effect observed in bacteria. Learn more about the different CRISPR applications in plants.     Transgenic ...helped us coordinate and promote the awards. References 1. Martinez-Klimova, Elena et al. “Endophytes...

  40. Management for Scientists: Managing vs. Leading

    Type
    Blog Post
    ...leader-specific. How is leadership different from good management? Aren't all career skills leaderhsip skills...because it emphasizes the importance of these very different roles. At any stage in our careers we may be in...you are not fulfilling the requirements of the current task. Most of us start as doers, some become managers...boss) that you are ready to try operating in a different role?  I think the secret sauce can be summed ...
Showing: 481 - 520 of 1018 results