Genomic DNA target sequence (eCas12f1-specific) (DNA target)

The target refers to a specific genomic DNA sequence recognized and cleaved by the engineered Cas12f1 (eCas12f1) system, a miniature CRISPR effector derived from Type V-F CRISPR-Cas systems (Kim et al., 2022, Nature Biotechnology). eCas12f1 is approximately one-third the size of Cas9, making it highly suitable for delivery using size-constrained vehicles like adeno-associated virus (AAV) for human gene therapy (Xu et al., 2021, Nature Communications). The target sequence must be located immediately adjacent to a T-rich protospacer adjacent motif (PAM), typically 5'-TTTR-3', which is essential for the initial binding and DNA unwinding by the eCas12f1-sgRNA complex (Karvelis et al., 2020, Nucleic Acids Research). Upon successful hybridization of the sgRNA spacer with the complementary DNA strand, the enzyme's RuvC nuclease domain generates a double-strand break (DSB). This targeted cleavage allows for precise genome modifications, including gene disruption via non-homologous end joining or gene correction via homology-directed repair, to treat various genetic disorders and cancers. The interaction between the eCas12f1-sgRNA complex and this DNA target is the fundamental mechanism for its application as a programmable genome-editing tool. Specificity is maintained through the complementarity between the sgRNA spacer and the target DNA, though potential off-target effects remain a primary safety concern in clinical applications.

Other names

CRISPR-eCas12f1 target siteProtospacerGenomic target locuseCas12f1-complementary DNA sequence

Site-specific DNA cleavage mediated by the RuvC nuclease domain of eCas12f1 following sgRNA-guided recognition of a T-rich protospacer adjacent motif (PAM) and hybridization with the complementary genomic DNA sequence.