...research. Image from Reproducibility for Everyone. Case study 1: Cell lines Many of the reagents used to...world shows that science is truly a global endeavor. Case study 2: Plasmids Another example of reagents that...Browse Addgene's COVID-19 plasmids and resources! Case study 3: Mouse strains Reagents developed during...

...unmeasurable skills and productivity coming from that casual discussion on the way to the office or in the kitchen... flexible work schedule the more we realize how casual chatter is very much missed in the virtual world...widely used and could bring back a taste of that casual kitchen chatter, ultimately sparking new ideas....

...Th2) cells from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation...Wang Lab created a tripartite system consisting of dCas9, a gRNA fused to an RNA binding motif, and a RNA-binding...protein-bEFP fusion. Thus, the bEFPs bind the UAS via the dCas9-gRNA complex which allows transcription from the...

...FAP - fluorogen activating protein - dL5** in this case), and 2) a fluorogen (the TAP - targeted and activated...activated photosensitizer - MG-2I in this case). The fluorogen is not fluorescent until it binds its protein...easy to watch development in real time.  In both cases, dL5** was tagged with mCerulean3 (mCer3) for visualization...

...in immunotherapy is the combination of the CRISPR/Cas9 methodology with T cell receptor engineering (Ref...using randomly integrating viral vectors, CRISPR/Cas9 genome editing allows targeted engineering of the.... "Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection." Nature 543.7643 (2017...

...depending on the vector and virus. Viral CRISPR-Cas9 tools have been engineered to introduce frameshift...be used to express proteins. Viral expression cassettes typically contain a promoter (inducible or constitutive... Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells ...

...analysis performed in experiment 1. Use camelCase: In camelCase, each subsequent word in a file name after... more than one word should be named in the camelCaseFormat since spaces between each word can make it ...

...' decisions to alter postdoc salaries - in some cases reversing these decisions entirely. At Future of...or are being compelled at the state level, in the case of some public institutions) to cancel plans to ...communication of their plans (many postdocs - and in some cases, faculty - have found out about what their institution...

...will be blocked during protein folding, in which case you should try tagging the opposite end.  If neither...disrupted by denaturants (Hochuli, 1988). In some cases, proteins can be bound under denaturing conditions... aren’t any good antibodies available. In these cases, having a tagged version of the protein is hugely...

...cytometry, secondary antibodies may be indicated on occasion, usually after other signal amplification techniques...indiscriminately, giving them the same output signal. In these cases, it is best to use the direct method when possible...confidently, collect a broader array of data, and occasionally ask themselves and others that most perplexing...

...entirely, and our software needs to handle these cases. The following code examples show how software can...sample and get it ready for shipping. In the first case, we naively assume that everything will go perfectly...we can interchange them easily as needed. In this case, the physical considerations of the laboratory make...

...single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how changing transfection...enough to get a sufficient titer; this is often the case with large genes. This is due to inefficient packaging...inserts; a study by Kumar et al. found that in some cases, titers can drop 10-fold for every 2kb increase ...

...those used for breeding heterozygous mice. In this case, your parental mouse should be hemizygous at the...similar epigenetic modifications. Special breeding cases Whether your mouse colony consists of genetically...explained more in Part 1 of this series). In any case, Cre is a dominant phenotype, while you’ll want ...

...your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene, we have the entire SapTrap toolkit.... "SapTrap, a toolkit for high-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans." Genetics...blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids 101 blog posts...

...other things, science communication comprises broadcasting, science writing, and certain aspects of art...audio or visual person. Why don’t you record a podcast or put some effort into making a video? Use your...Communication Posts Browse our Career Page Listen to Our Podcast Episode on Science Communication Additional Resources...

...they’re knowingly selling a bad product (in which case, you should probably stop using that company). Frequently...frequently asked by customers (and I expect this is the case at most companies). Always go back to the source...that could take weeks to resolve adequately. Occasionally there are some really off-the-wall questions...

...always maintain a backup replicates of each clone in case of contamination. We do not normally use antibiotics...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...potentially large number of single clones obtained (on occasion we have prepared two 48-well plates of clones)...

...endoplasmic reticulum localization, Munro and Pelham casually introduced the Myc-tag (Munro, et al. 1986; Munro...developed almost 40 years ago? Well…kinda. In some cases, antibody providers are distributing monoclonal ...same hybridoma developed by Evans, et al. In other cases, providers have moved to producing anti-Myc [9E10...

... matches the copy number of your sample. In that case, an RQ value of one (or, more realistically, between...ddPCR may be a better option for more challenging cases.  Price point If budget is a concern, look for...number and it may not be able to identify trends in cases where the copy number variation is two or less (...

...individual expression cassettes, having the genes expressed from the same cassette is sometimes advantageous...and you can then insert the whole multicistronic cassette into a backbone as a single unit. Although it ...