CRISPRbased

CRISPRbased refers to technologies that employ CRISPR systems to alter genetic material, regulate gene expression, or detect nucleic acids. Core components typically include a CRISPR-associated nuclease and a programmable guide RNA that directs the enzyme to a complementary target sequence adjacent to a protospacer adjacent motif (PAM). In the widely used CRISPR-Cas9 system, the nuclease creates a double-strand break, which the cell repairs via non-homologous end joining or homology-directed repair, enabling gene disruption or targeted sequence replacement. Catalytically inactive variants (dCas9) serve as DNA-binding scaffolds for transcriptional regulation or epigenetic editing.

Other CRISPR effector proteins such as Cas12a and Cas13 broaden the scope to different nucleic acids and offer alternative PAM requirements and diagnostic utilities. Cas12a cleaves DNA with staggered ends and exhibits collateral single-stranded DNA activity, while Cas13 targets RNA and can show collateral RNase activity, useful in certain detection platforms.

Applications span basic research, functional genomics, and prospective therapeutics. Base editing and prime editing enable precise base substitutions or targeted sequence changes without introducing double-strand breaks. CRISPR-based diagnostics use collateral cleavage to detect specific genetic sequences, with platforms such as SHERLOCK and DETECTR.

Delivery and safety considerations include methods like viral vectors and non-viral systems, tissue specificity, editing efficiency, and potential off-target effects. Immunogenicity of Cas proteins, mosaic edits in multicellular organisms, and ethical or regulatory issues, particularly surrounding germline edits, are active areas of discussion. Ongoing research aims to improve specificity, broaden PAM compatibility, and develop safer, more effective delivery approaches.