Preprint led by @ruth_e_hanna on using base editors for large-scale screens in human cells. A thread. A long thread. I'm sorry :) biorxiv.org/cgi/content/sh…
Figuring out what genes do is hard, figuring out individual nucleotides even harder, so we wanted to see if C>T base editors were up to the task in pooled screens. First, benchmarking using a tiling library where loss-of-function mutations will have a known phenotype.
Aug 28, 2019 • 14 tweets • 13 min read
New @biorxivpreprint from the group, setting our sites on Cas12a (the #CRISPR enzyme formerly known as Cpf1) as a simpler system for multiplexed genetic screens. Thread...
Optimization of AsCas12a for combinatorial genetic screens in human cells biorxiv.org/content/10.110…@biorxivpreprint Cas12a has a major benefit, only needing one promoter for expressing multiple guides, and has a smaller constant region (the tracrRNA-like ‘handle’ for Cas12a, the DR, is only 20nts). So, a second guide is only 43 more nts -- much easier to write (synthesize) and read (sequence)
May 31, 2018 • 11 tweets • 2 min read
Useful paper, comparing multiple Cas proteins in mammalian cells. A few thoughts... (thread) genomebiology.biomedcentral.com/articles/10.11…
First, a word of caution on generalizability. All Cas enzymes are expressed with the same NLS / tag arrangements. Totally obvious, experimentally, why this was done. However, can't assume they do (or don't) interfere with function equally.
