Spacers/protospacers In a bacterial genome, CRISPR loci contain "spacers" (viral DNA inserted into a CRISPR locus) that in type II adaptive immune systems were created from invading viral or plasmid DNA (called "protospacers") Upon subsequent invasion, a CRISPRassociated nuclease such as Cas9 attaches to a tracrRNA–crRNA complex, which guides Cas9 to the invadingCRISPRDT (CRISPR DNA Targeting) is a web application to help biologists design optimal gRNAs for the CRISPRCpf1 system CRISPRDT is essentially composed of many interfaces and a backend pipeline Maximum number of mismatches and gaps in seed region tolerated by off targets The seed region stands for 6 nucleotides in the 5' PAMRecently, Wessels et al showed that the seed region (15–21 nucleotides) in gRNA is critical for CRISPR/Cas13d activity A single mismatch in the seed region led to diminished activity, and two or three mismatches led to severe reduction in Cas13d activity in this study
High Fidelity Crispr Cas9 Increases Precise Monoallelic And Biallelic Editing Events In Primordial Germ Cells Engormix
