2023
DOI: 10.1016/j.cej.2023.144407
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Branched DNA switchable CRISPR-Cas12a system for sensing FEN1 activity

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Cited by 12 publications
(5 citation statements)
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“…The reason was that the nick outside the seed region could not share the benefits of Cas12a’s capacity to enhance the combination of nicked TS (9–15 nt) and crRNA. As a result, the binding affinity between nicked TS and crRNA was insufficient, which affected the formation of the R-loop and adequately attenuated the cis/trans -cleavage activity of Cas12a, which was consistent with previously published data. , Since dsDNA with 17 bp could activate Cas12a by itself, nick located at 17 or 19 nt from PAM had no impact on Cas12a . From the above results, it could be confirmed that the DREAM system was also intolerable to the nicks in the middle of the protospacer sequence, which was similar to the results of the AP site modification.…”
Section: Results and Discussionsupporting
confidence: 90%
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“…The reason was that the nick outside the seed region could not share the benefits of Cas12a’s capacity to enhance the combination of nicked TS (9–15 nt) and crRNA. As a result, the binding affinity between nicked TS and crRNA was insufficient, which affected the formation of the R-loop and adequately attenuated the cis/trans -cleavage activity of Cas12a, which was consistent with previously published data. , Since dsDNA with 17 bp could activate Cas12a by itself, nick located at 17 or 19 nt from PAM had no impact on Cas12a . From the above results, it could be confirmed that the DREAM system was also intolerable to the nicks in the middle of the protospacer sequence, which was similar to the results of the AP site modification.…”
Section: Results and Discussionsupporting
confidence: 90%
“…It is hypothesized that the reason has to do with Cas12a's greater tolerance to ssDNA. 10,24 In conclusion, it could be confirmed that the DREAM system had a high tolerance to the AP sites at the 5′ or 3′ end of the protospacer sequence while the intermediate modification of the AP site was intolerable.…”
Section: ■ Introductionmentioning
confidence: 73%
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“…The equation for the regression analysis was determined as y = 265.8438 x + 455.5189, with a calculated limit of detection of ATP as 0.046 μM. In order to ascertain the continued high sensitivity of our biosensor in detecting intricate samples, 35–37 we employed the biosensor to assess distinct ATP concentrations present in 10% serum. In accordance with the observations portrayed in Fig.…”
Section: Resultsmentioning
confidence: 99%
“…The recent discovery of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems, especially the type V and VI Cas effectors (Cas12a, Cas13a, and Cas14), have introduced a new biosensing system for nucleic acid detection. For example, the CRISPR/Cas13a system, possessing high-efficient trans -cleavage activity activated by crRNA-guided cis recognition, can directly sensing of RNA targets at sub-pM concentrations . For highly sensitive applications, this CRISPR / Cas13a system often requires the combination of target amplification through PCR and other isothermal amplification technologies. Although the introduction of preamplification could attain extensive enhancement in assay sensitivity and specificity, the preamplification step also might increase the possibility of aerosol contamination and complex the analytical procedure.…”
mentioning
confidence: 99%