CRISPR-Cas9 History and Engineering
A modular platform using engineered single short guide RNA to allow programming of CRISPR specificity, permitting high efficacy gene induction for analysis of gene function.
The ability to control gene expression has been the key method in elucidating their respective functions, pathways, and regulatory elements; paving a way for future therapeutic applications.
The two main approaches of determining gene function involve the analysis of loss-of-function (LOF) and gain-of-function (GOF) mutations. LOF involves a mutation in an allele where partial or full loss in genetic function occurs. GOF involves the introduction of a mutation which generates a new allele associated with a new function. The problem with GOF screening approaches is that theyre hindered by a requirement for large comprehensive cDNA library overexpression systems which rarely encompass the full spectrum of isoform variation. Viral expression vectors are not large enough to allow these to be cloned.
