...AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal cord...be identified by PCR later. The PNS presents a different problem: it’s spread throughout the body and is... and supplemental figure 8 in Chan et al).  References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered...selection yields AAV variants for widespread gene transfer to the adult brain. Nature biotechnology. 2016...

...more flexible than Cas9/Cpf1. Some Cas13 enzymes prefer targets with a given single base protospacer flanking...are present in the gRNA spacer, as ADAR will preferentially edit an adenine if the template has a cytosine...that a S375A mutation in RESCUE (named RESCUE-S) offered ~76% on-target C-to-U editing while reducing off-target... Valley contributed to updating this article. References Cox, David B.T., et al. “RNA editing with CRISPR-Cas13...

...the RPE65−/− dog, was characterized and found to suffer from early and severe visual impairment that faithfully...falling from grace, gene therapy is back, better and safer. It is now one of the most active and exciting areas...and the development of personalized medicine. References and further reading 1. FDA approves gene therapy.... PubMed PMID: 29983335. 9. Day TP1, Byrne LC, Schaffer DV, Flannery JG, 2014. Advances in AAV vector ...

...park. Happy editing!   Resources and References References Yu, Y., Guo, Y., Tian, Qigi, Lan, Y., et...the break, potentially shuttling repair down a different pathway (Charpentier et al., Nat Com).   ...2014) 10.7554/eLife.04766 Moreno-Mateos, M. A., Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1...

...While I can’t make it “all better” for you, I can offer some tips and tactics that have worked for my friends...miserable at home or work, your relationships will suffer in both the short run and the long run and so will...alone, it should be obvious that your career will suffer. And, yes, your partner CAN take the kids to the... or her own dinner if you are going away to a conference. Really. No one will starve while you are away...

... optical pooled screens in human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin...editing efficiencies in a number of cell types at different genomic loci — in some cases, over 20-fold higher...prime editor. B, Prime editing efficiencies at different loci in U2OS cells. Image adapted from Yan et ...that an A122D mutation in the rhodopsin domain conferred several new improvements. The new variant has ...

...if your model is correct or your data favor a different model. Finding evidence for or against a model.../goal, an approach, anticipated outcomes, and preferably a description of the payoff if the aim is successful...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

... studies.     Jennifer Tsang contributed to updating this article. References Abudayyeh OO, Gootenberg...variations of Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition...Zetsche et al., 2015 ). Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends...

...immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, packaging...equally potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs...Sontheimer lab used NmeCas9 with AAV to edit two different disease-causing loci in mouse liver.  Two other...have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW, Mali P, Wu...

...can be used with or as fluorophores in several different ways. Researchers have used oligonucleotides in...biosensors. Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied and used in...were born. Aptabeacons can be created in many different ways, but the general premise is that an aptamer...Aptamers can be selected for these small ligands. References 1. Ma, Hanhui, et al. “Multicolor CRISPR labeling...

...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi E,...RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using Cas7-11   Use ...fibrosis, which can be caused by thousands of different mutations, all found in the same gene. If we only...

...Addgenie Hannah Dotson.  Resources and references References Genome-scale CRISPR-Cas9 knockout and transcriptional...properly amplify.  Amplification protocols therefore differ from library to library. We strongly recommend ...Addgene’s pooled libraries use these protocols.  References McDade JR, Waxmonsky NC, Swanson LE, Fan M. ...

...options in our CRISPR 101 series!  References and resources References  Anzalone, A. V., Koblan, L. W. &...plasmids that have been optimized for expression in different species and cell types including (but not limited...101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled Library...

... The entry clone and destination vector carry different antibiotic resistance markers (indicated here ...STEP 1: Generate an Entry Clone There are a few different ways to generate our desired entry clone - human...interest: Find Gateway cloning plasmids here! References 1. Chee JY, Chin CF (2015) Gateway Cloning Technology... Digest Video Check Out Our Molecular Biology Reference Pages  ...

...that “… the emergence of the video abstract genre offers a low-barrier opportunity for researchers to leverage...to measure the impact of their contribution. It offers a 5-step process and several examples for creating...professors at the University of Washington in St. Louis, offer guidance on how to make a low-cost whiteboard video...WeShareScience or email Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric...

...of conversations with people who work in many different scientific fields. They also host AMA forums where...team with a description of our AMA and Joanne’s preferred time to start answering questions. r/Science has...AMA is now a great resource for any scientists to refer to if they have questions about science careers,...future. The AMA was also cataloged for permanent reference by the Winnower. The only small downside to the...

...Vader in "Return of the Jedi" While Vader was referring to the evil side of a mystical "Force," this quote...cannot be ignored. They have the potential to interfere with the function of resident ER chaperones and...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...Kaposi’s sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent...

...of veracity of the conclusions drawn. The main difference with the PLOS policy is that access of this data...perhaps similarly impoverished lab) might apply different or novel tools and assumptions to reveal findings...geneticists may track the motion of multiple animals on different occasions via video, generating huge amounts of...responsible for storage.  If datasets are too large to transfer to a journal repository, they are likely too large...

...to a mouse model of muscular dystrophy. Three different splice site donor and acceptor sequences were ...reconstituted after integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains...greater and more consistent transgene expression. References: Allocca M, Doria M, Petrillo M, Colella P, Garcia-Hoyos...retinal gene therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out ...

...flanking a FLOXed neoR gene. pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR...Schofield, 2015; Moser and Hirsch, 2016; Epstein and Schaffer, 2017) Hopefully you’ve been able to check out...cohesin gene inactivation in human cancer.     References 1. Russell, David W., and Roll K. Hirata. "Human...27280971. 10. Epstein, Benjamin E., and David V. Schaffer. "Combining engineered nucleases with adeno-associated...