In this review, we aim to provide a mechanistic insight into the working mechanism of Cas12a, comparing it with Cas9, and eventually provide an overview of its current applications in genome editing and biotechnology applications.

In this review, we aim to provide a mechanistic insight into the working mechanism of Cas12a, comparing it with Cas9, and eventually provide an overview of its current applications in genome editing and biotechnology applications.

2024年12月4日 · CRISPR-associated Cas12 proteins are a highly variable collection of nucleic acid-targeting proteins. All Cas12 variants use RNA guides and a single nuclease domain to target complementary DNA or,...

2023年9月5日 · Cas12a, a CRISPR-associated protein complex, has an inherent ability to cleave DNA substrates and is utilized in diagnostic tools to identify DNA molecules. We demonstrate that multiple...

2024年7月25日 · Cas12a scans DNA for a PAM sequence with a highly flexible REC1 domain. PAM recognition leads to local DNA melting and seed formation. Cas12a continues to check for guide-target complementarity during R-loop extension and limited R-loop contacts ensure an unstable and readily reversible R-loop.

4 天之前 · For transduction of Cas12a-expressing lentiviral vectors, approximately 4.0 × 10 6 cells were seeded and transduced with Cas12a-encoding lentiviral vectors at a MOI of 1 in the presence of 10 μg ...

In its wake, the novel RNA-guided endonuclease system CRISPR-Cas12a is transforming biological research due to its unique properties, such as its high specificity or its ability to target T-rich motifs, to induce staggered double-strand breaks and to process RNA arrays.

2024年8月25日 · We reveal that the activated crRNA–Cas12a RNP favors the trans-cleavage of longer sequences than commonly used. These new findings of the RNA-activated trans -cleavage capability of Cas12a provided the foundation for the design and construction of CRISPR nanorobots that operate in living cells.

2025年3月19日 · The formed duplex, in conjunction with the crRNA-handle, activates Cas12a's trans-cleavage activity. This leads to cleavage of a fluorescent reporter, generating an enhanced signal. The method enables direct RNA detection without reverse transcription or sample amplification, offering simplicity and efficiency. This method demonstrates high ...

3 天之前 · The PCR-coupled CRISPR/Cas12a-based fluorescence assay includes (i) a pre-amplification step and (ii) a nucleic acid detection step. The pre-amplification step is based on a commonly used PCR, and the detection step is based on the CRISPR/Cas12a property to nonspecifically hydrolyze single-stranded DNA fluorescent reporter molecules.