Off-target genomic DNA loci with partial complementarity to the MACF1 guide RNA (MACF1 off-target sites)

Off-target genomic DNA loci with partial complementarity to the MACF1 guide RNA represent unintended sites within the genome where gene-editing machinery, such as CRISPR-Cas9, may bind and introduce modifications (Fu et al., 2013, Nature Biotechnology). These sites possess sequence homology to the primary target sequence within the Microtubule-actin cross-linking factor 1 (MACF1) gene, which is a large cytoskeletal protein involved in linking actin filaments and microtubules to regulate cell migration and signaling (Hu et al., 2017, Nature Communications). The interaction at these off-target loci is a major safety concern in the development of genetic therapies, as it can lead to permanent and potentially deleterious mutations (Zhang et al., 2015, Science). Such unintended edits may result in the disruption of essential genes, chromosomal rearrangements, or the activation of oncogenes, posing a risk of genotoxicity or malignant transformation (Frock et al., 2015, Nature Biotechnology). Consequently, rigorous screening using high-throughput sequencing methods like GUIDE-seq or CIRCLE-seq is required to identify these sites and optimize guide RNA specificity (Tsai et al., 2015, Nature Biotechnology). Understanding and mitigating these off-target effects is essential for ensuring the precision and safety of any therapeutic intervention targeting the MACF1 locus.

Other names

MACF1 off-target sequencesNon-specific MACF1 gRNA binding sitesUnintended MACF1 CRISPR cleavage sitesMACF1 gRNA mismatches

Unintended DNA cleavage and subsequent repair (via Non-Homologous End Joining or Homology-Directed Repair) at genomic locations that share sequence similarity with the intended MACF1 target sequence (Hsu et al., 2013, Nature Biotechnology).