Mar 12, 2018 · After careful analyses of experimental conditions, we found that only the ternary complex of Cas12a/crRNA/targeted ssDNA (or targeted dsDNA) was able to cleave the 5′-labelled target ssDNA...

Feb 15, 2018 · Here we show that RNA-guided DNA binding unleashes indiscriminate single-stranded DNA (ssDNA) cleavage activity by Cas12a that completely degrades ssDNA molecules. We find that target-activated, nonspecific single-stranded deoxyribonuclease (ssDNase) cleavage is also a property of other type V CRISPR-Cas12 enzymes.

Besides high-specific dsDNA cleavage, Cas12a has also been shown to exhibit indiscriminate ssDNA degradation activity upon activation with a ssDNA complementary to the crRNA guide. This activity is displayed by all Cas12a orthologs and degrades any available ssDNA molecule into single/double nucleotides [ 40 ].

Sep 27, 2024 · Among the discovered CRISPR-Cas enzymes, CRISPR-Cas12a exhibits a unique ability that can indiscriminately cut any nearby non-specific single-stranded DNA (ssDNA) molecules after...

Here we show that RNA-guided DNA binding unleashes robust, indiscriminate single-stranded DNA (ssDNA) cleavage activity in Cas12a sufficient to completely degrade both linear and circular ssDNA molecules.

Apr 27, 2018 · CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems. Like CRISPR-Cas9, Cas12a has been harnessed for genome editing on the basis of its ability …

Mar 5, 2024 · Control of CRISPR/Cas12a trans -cleavage is crucial for biosensor development. Here, we show that small circular DNA nanostructures which partially match guide RNA sequences only minimally...

Jun 24, 2022 · CRISPR-Cas12a (Cpf1) is a bacterial RNA-guided nuclease that cuts double-stranded DNA (dsDNA) at sites specified by a CRISPR RNA (crRNA) guide. Additional activities have been ascribed to this enzyme in vitro: site-specific (cis) single-stranded DNA (ssDNA) cleavage and indiscriminate (trans) degrad …

Dec 13, 2018 · Single-molecule FRET provides the thermodynamics and kinetics of the conformational activation leading to phosphodiester bond hydrolysis. These findings illustrate why Cas12a cuts its target DNA and unleashes unspecific cleavage activity, degrading ssDNA molecules after activation.

Mar 26, 2025 · Upon recognition by the Cas12a/gRNA complex, the ssDNA-FQ reporter is cleaved, producing a strong fluorescent signal (output = 1). ... CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems. Like CRISPR-Cas9, Cas12a has been harnessed …