Genomic DNA at guide-RNA–specified target sequence (gRNA-targeted DNA)

Genomic DNA at a guide-RNA–specified target sequence refers to the specific chromosomal locus recognized and bound by a CRISPR-Cas ribonucleoprotein (RNP) complex for therapeutic modification. This targeting is mediated by Watson-Crick base pairing between a synthetic guide RNA (gRNA) and a complementary DNA sequence, typically requiring a nearby protospacer adjacent motif (PAM) for enzyme recognition (Jinek et al., Science, 2012). Once localized, the associated nuclease (such as Cas9 or Cas12a) induces a double-strand break or performs precise chemical base modifications. These events trigger cellular repair pathways like non-homologous end joining (NHEJ) or homology-directed repair (HDR), allowing for the permanent disruption of disease-causing genes or the correction of mutations (Doudna & Charpentier, Science, 2014). This target is the foundation of the first FDA-approved CRISPR therapy, exagamglogene autotemcel, which targets the BCL11A enhancer to treat hematological disorders (FDA, 2023). While highly effective for precision medicine, therapeutic use requires rigorous monitoring for off-target activity and potential long-term genotoxicity (Fu et al., Nature Biotechnology, 2013).

Other names

CRISPR target siteProtospacerGenomic target locusgRNA-directed DNA sequenceCRISPR-Cas9 target sequence

Site-specific DNA modification via CRISPR-Cas mediated cleavage, base editing, or prime editing to disrupt, correct, or regulate gene expression at a precise genomic location.