Title : CRISPR designing your G-rna for CAS 9 genome editing using DNA sensors to detect cerebrospinal fluid morbiditidity in-vitro viral gene transmission
CRISPR technology has made it easier than ever both to engineer specific DNA edits and to perform functional screens to identify genes involved in a phenotype interest.
We will discuss differences between these approaches, and provide updates on how best todesign gRNAs. The hammer, the jigsaw, and the wrench are all great tools, but which one you use, of course, depends on what you are trying to do – there’s no “best” tool among them. While this seems obvious, it is important to remember that the same is true when designing gRNAs for using CRISPR technology – the “best” gRNA depends an awful lot on what you are trying to do: gene knockout, a specific base edit, or modulation of gene expression. Location and sequence are important considerations for designing your gRNAs. For indels, it's not so important what location in the gene you target, but it is important that your gRNA sequence is designed to be highly active and reduce off targets. For CRISPRa and CRISPRi, these considerations are of roughly equal importance (target should be near the TSS but you can worry less about sequence optimality because you generally have fewer sequences to choose from). Finally, for HDR, location is much more important because you have to target within ~30 nt of your proposed edit, which means there are so few gRNAs to choose from that sequence preferences must largely be ignored.