CRISPR –Cas9 gene editing induced complex on-target outcomes in human cells

Clustered, regularly interspaced, short palindromic repeats (CRISPR)-Cas9 has been a promising tool for gene engineering, such as correcting disease-associated mutant alleles in somatic or stem cells [1]. Cas9 is a single endonuclease evolved in bacteria and archaea to function as a natural adaptive immune system [2, 3]. Cas9 programmed with crRNA (CRISPR RNA) and tracrRNA (trans-activating crRNA) (Cas9-sgRNA ribonucleoprotein complex) has HNH and RuvC nuclease domains to cleave target DNA, generating two blunt ends of double-strand breaks (DSBs), usually three bp upstream of a protospacer adjacent motif (PAM, NGG for SpCas9 from Streptococcus pyogenes) sequence [3, 4].

Source: Experimental Hematology - March 15, 2022 Category: Hematology Authors: Wei Wen, Xiao-Bing Zhang Source Type: research