Uracil-Mediated New Photospacer-Adjacent Motif of Cas12a To Realize Visualized DNA Detection at the Single-Copy Level Free from Contamination
The CRISPR/Cas12a (cpf1) system was reported to indiscriminately cleave single-stranded DNA after binding with target DNA strands. This usually required the target DNA strands to contain the protospacer-adjacent motif (PAM) sequence of TTTN. Herein, we found Cas12a can also recognize another PAM seq...
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| Vydané v: | Analytical chemistry (Washington) Ročník 91; číslo 17; s. 11362 |
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| Hlavní autori: | , , , , , |
| Médium: | Journal Article |
| Jazyk: | English |
| Vydavateľské údaje: |
United States
03.09.2019
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| ISSN: | 1520-6882, 1520-6882 |
| On-line prístup: | Zistit podrobnosti o prístupe |
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| Shrnutí: | The CRISPR/Cas12a (cpf1) system was reported to indiscriminately cleave single-stranded DNA after binding with target DNA strands. This usually required the target DNA strands to contain the protospacer-adjacent motif (PAM) sequence of TTTN. Herein, we found Cas12a can also recognize another PAM sequence of UUUN resulting in activation of its ssDNA collateral cleavage effect. To make this finding useful, by combining with LAMP, we first realized CRISPR/Cas12a for directly visualized DNA detection at the single-copy level. By treating with UDG enzyme, we made this system free from residual amplicon contamination, which is a big problem in this field. Thus, an ultrasensitive and anticontaminant DNA detection platform, namely, UDG and LAMP and CRISPR (ULC). This new finding would help us better understand the mechanism of Cas12a and expand its application. |
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| Bibliografia: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| ISSN: | 1520-6882 1520-6882 |
| DOI: | 10.1021/acs.analchem.9b02554 |