Genomic DNA at single-guide RNA-specified loci (gDNA (sgRNA-specified))

Genomic DNA at single-guide RNA (sgRNA)-specified loci serves as the fundamental target for CRISPR-Cas9 and related genome-editing therapeutic platforms. In this context, the target is a specific sequence of nucleotides within the human genome that is complementary to a synthetic sgRNA, which directs a Cas nuclease to induce a site-specific modification [Jinek et al., 2012, Science]. This interaction allows for the precise disruption of disease-causing genes, the correction of point mutations, or the targeted insertion of therapeutic transgenes [Doudna & Charpentier, 2014, Science]. For instance, the FDA-approved therapy exagamglogene autotemcel targets the BCL11A erythroid-specific enhancer to treat sickle cell disease by inducing fetal hemoglobin expression [FDA, 2023, Casgevy Prescribing Information]. Beyond monogenic disorders, this target is being explored for treating infectious diseases and cancer through the modification of immune cells or viral DNA [Gillmore et al., 2021, NEJM]. The therapeutic utility of these loci depends on the specificity of the sgRNA-DNA binding, as unintended interactions at off-target sites can lead to genomic instability or oncogenic transformations [Fu et al., 2013, Nature Biotechnology]. Consequently, the characterization of these loci involves extensive bioinformatic prediction and empirical validation to ensure safety and efficacy in clinical applications [Tsai et al., 2015, Nature Methods].

Other names

CRISPR target siteProtospacersgRNA-targeted DNAGenomic target locusRNA-guided DNA target

Site-specific DNA modification via RNA-guided endonucleases (e.g., Cas9, Cas12a) followed by cellular DNA repair mechanisms like Non-Homologous End Joining (NHEJ) or Homology-Directed Repair (HDR) [Jinek et al., 2012, Science].