Off-target genomic loci of Actin alpha 2 (ACTA2)-directed single guide RNA (sgRNA) (ACTA2 sgRNA off-targets)

Off-target genomic loci of ACTA2-directed sgRNA refer to unintended DNA sequences that share partial homology with the single guide RNA (sgRNA) designed to target the ACTA2 gene. ACTA2, which encodes alpha-smooth muscle actin, is a critical gene involved in vascular contractility and is often targeted in research related to thoracic aortic aneurysms and fibrotic diseases (UniProt P62736). During CRISPR-Cas9 mediated editing, the Cas9 enzyme may recognize and cleave these off-target sites due to sequence similarity, leading to permanent mutations such as insertions or deletions (PubMed 25513782). These genomic alterations pose significant safety risks, including the potential for insertional mutagenesis or the disruption of essential regulatory elements (PubMed 28504668). In the context of therapeutic development, identifying and minimizing these off-target effects is crucial to prevent adverse outcomes such as oncogenesis or impaired cellular function (PubMed 19409525). Monitoring these sites typically involves high-throughput sequencing methods like GUIDE-seq and CIRCLE-seq to ensure the specificity and safety of the genetic intervention (PubMed 28504668).

Other names

ACTA2 CRISPR off-targetsUnintended Cas9 cleavage sitesHomologous genomic sequencesNon-specific sgRNA binding sites

Unintended DNA cleavage and subsequent repair via non-homologous end joining (NHEJ) or homology-directed repair (HDR) at genomic sites with sequence similarity to the ACTA2 target sequence.