Direct observation of the formation of a CRISPR-Cas12a R-loop complex at the single-molecule level
Here, we develop an optical tweezers-based single-molecule manipulation assay to detect the formation of an R-loop complex in the Cas12a system and characterize its thermodynamic stability. We found that the formation of the R-loop complex induces a two-step unfolding of a DNA hairpin containing the...
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| Veröffentlicht in: | Chemical communications (Cambridge, England) England), 2020-02, Vol.56 (14), p.2123-2126 |
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| container_title | Chemical communications (Cambridge, England) |
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| creator | Cui, Yang Tang, Yangchao Liang, Meng Ji, Qinghua Zeng, Yan Chen, Hui Lan, Jie Jin, Peng Wang, Lei Song, Guangtao Lou, Jizhong |
| description | Here, we develop an optical tweezers-based single-molecule manipulation assay to detect the formation of an R-loop complex in the Cas12a system and characterize its thermodynamic stability. We found that the formation of the R-loop complex induces a two-step unfolding of a DNA hairpin containing the target sequence, the non-target sequence binds loosely to Cas12a and can be easily released from the complex, and the Nuc domain of Cas12a plays key roles in target binding and R-loop formation.
An optical tweezers-based single-molecule manipulation assay was developed to detect the formation of an R-loop complex in the CRISPR-Cas12a system. |
| doi_str_mv | 10.1039/c9cc08325a |
| format | Article |
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An optical tweezers-based single-molecule manipulation assay was developed to detect the formation of an R-loop complex in the CRISPR-Cas12a system.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>31970368</pmid><doi>10.1039/c9cc08325a</doi><tpages>4</tpages><orcidid>https://orcid.org/0000-0003-4031-6125</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1359-7345 |
| ispartof | Chemical communications (Cambridge, England), 2020-02, Vol.56 (14), p.2123-2126 |
| issn | 1359-7345 1364-548X |
| language | eng |
| recordid | cdi_pubmed_primary_31970368 |
| source | MEDLINE; Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
| subjects | CRISPR-Cas Systems - genetics DNA - chemical synthesis DNA - chemistry DNA - genetics Optical Tweezers Thermodynamics |
| title | Direct observation of the formation of a CRISPR-Cas12a R-loop complex at the single-molecule level |
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