...The lab additionally recommends using an anti-mCherry antibody rather than endogenous mCherry fluorescence...transduction with this virus. While various Cas9 mutants have been engineered to perform unique editing ...

... 1990’s, microRNAs (miRNAs) are recognized as important regulators of post-transcriptional gene expression...in vivo, the non-immunogenic violet-excited GFP variant Vex. To understand the impact of this gene editing...immune response other than what would be the anticipated effect of the targeted gene. CHIME can also be...

...handle, but I wasn’t using them to my professional advantage. It wasn’t until I wrote a blog post about a research...start using social media to their professional advantage, I’ve selected five resources that have tips for...can fit into a researcher’s work day. It’s also important to remember that it’s not the amount of time you...

...updated Jul 20, 2021. By Will Arnold and Shreya Vedantam Accelerating research by improving the availability...expression vectors containing various spike protein variants are also available. Plasmids are continually added...these plasmids available for scientists speedily meant that we had to revamp some of our quality control...

...cloning: A SLIC makeover In 2007, LIC received an important update, courtesy of Addgene depositor Stephen ...structures at the ends of fragments. The major advantage of SLIC over Gibson assembly is cost, as T4 polymerase...homologous recombination in vitro to generate recombinant DNA via SLIC. Li MZ, Elledge SJ. Nat Methods....

... virus is produced in just a few days. Another variant, Phoenix-gp, contains only Gag-Pol and enables ...γ-retroviruses differ from lentiviruses? There are a few important differences between engineered γ-retroviruses ...names? The nomenclature of the plasmid backbone is meant to be descriptive of the components found within...

...allows you to infect cells and deliver cargo, it’s important to understand their differences to choose the ...used to achieve transient, high expression of the relevant cargo without the fear of random genomic integration...non-integrating and infect non-dividing cells. AAVs primary advantage over adenovirus is their low immunogenicity, which...

...polymerase. E4/E1A activate transcription of other relevant viral genes (including the ones listed previously... it to evade detection by the host. Of note, recombinant adenovirus is missing the endogenous E1 gene,...services for some of the plasmids in our collection. Want to make a vector construct on your own? No problem...

...may seem tedious at first, especially since you’ll want to dig in to the computation, but they will serve...specific goal for each analysis will help you locate relevant information in the future. Approach: Outline...can store any power point, excel, or other files relevant to the project. README: The README.txt document...

...  and that’s just to name a few. But what if you want to study RNA-protein interactions? The characterization...RNAs and proteins with UV light, followed by an antibody pulldown of a protein of interest, much like using...disease. RaPID can also be used to test how RNA variants associated with human disease affect protein binding...

...riboprobe binding (via hybridization) specific antibodies that detect the mRNA-riboprobe complex are added...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...Addgene. If you use these published protocols and want to share pictures, feedback etc. please write to...

... deliver Cas9 variants, such as a transcriptional activator dCas9-VPR, if these variants are fused to ...Mature Nanoblades then bud into the cell culture supernatant and can be purified by centrifugation. Gag-Cas9...

...publication.   3’UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans...Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans...lab in 2015 adapted the AID system discovered in plants to conditionally deplete proteins of interest in...

...active in vitro and in human and plant cells. Its small size offers advantages for cellular delivery. Get ...integration and connectivity without the constraints of antibody labeling. This opens the door to in vivo experiments...

...where you want it to? Knowing that a gene is expressed in a particular cell type is important not only ...Gross-Scherf, B., Morikawa, R.K., Rompani, S.B., Hantz, P., Szikra, T., Esposti, F., Cowan, C.S., Bharioke...

...scientific conference or a karaoke bar), you probably want to sigh. The problem is that many of these studies...Sapozhnikov and Szyf 2021, Charlton et al. 2020). Most importantly, TET proteins affect transcription independent... directly inside the gRNA sequence is a great guarantee that DNMT1 will be blocked (within PAM works, ...

... Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in...2015). PubMed PMID: 26422227.   Potential advantages of Cpf1 Type II CRISPR systems based on Cas9 ...efficiency across CRISPR nucleases and NgAgo is an important next step for the field. Based on what we know...

...poster you want to know more about? Interested in doing a postdoc in a presenter’s lab? Want to collaborate...breaths the day-of, you’ll handle them with ease. Anticipate questions: When practicing with your colleagues...

...with all these CRISPR plasmids (and libraries)? We wanted to know, so we classified CRISPR plasmids containing...other countries, from Japan to Australia, are significant depositors and requestors. CRISPR tools are developed...our CRISPR depositors and requestors. Thank you!  Want to dive deeper into Addgene’s CRISPR data? Read ...

...difficult” GC-rich regions of the genome, which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and...researchers were unhappy with the new method and 65% wanted to wait until someone else optimizes an NgAgo protocol...wouldn’t be a surprise if someone came up with a mutant NgAgo (or a novel endonuclease) protein that can...