Miniature CRISPR-Cas12f1-Mediated Single-Nucleotide Microbial Genome Editing Using 3 '-Truncated sgRNA
DC Field | Value | Language |
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dc.contributor.author | Lee, Ho Joung | - |
dc.contributor.author | Kim, Hyun Ju | - |
dc.contributor.author | Lee, Sang Jun | - |
dc.date.accessioned | 2023-02-15T05:41:34Z | - |
dc.date.available | 2023-02-15T05:41:34Z | - |
dc.date.issued | 2023-02 | - |
dc.identifier.issn | 2573-1599 | - |
dc.identifier.issn | 2573-1602 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/60596 | - |
dc.description.abstract | The CRISPR-Cas system has been used as a convenient tool for genome editing because the nuclease that cuts the target DNA and the guide RNA that recognizes the target are separated into modules. Cas12f1, which has a smaller size than that of other Cas nucleases, is easily loaded into vectors and is emerging as a new genome editing tool. In this study, AsCas12f1 was used to negatively select only Escherichia coli cells obtained by oligonucleotide-directed genome editing. Although double-, triple-, and quadruple-base substitutions were accurately and efficiently performed in the genome, the performance of single-base editing was poor. To resolve this limitation, we serially truncated the 3 '-end of sgRNAs and determined the maximal truncation required to maintain the target DNA cleavage activity of Cas12f1. Negative selection of single-nucleotide-edited cells was efficiently performed with the maximally 3 '-truncated sgRNA-Cas12f1 complex in vivo. Moreover, Sanger sequencing showed that the accuracy of single-nucleotide substitution, insertion, and deletion in the microbial genome was improved. These results demonstrated that a truncated sgRNA approach could be widely used for accurate CRISPR-mediated genome editing. | - |
dc.format.extent | 10 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | MARY ANN LIEBERT, INC | - |
dc.title | Miniature CRISPR-Cas12f1-Mediated Single-Nucleotide Microbial Genome Editing Using 3 '-Truncated sgRNA | - |
dc.type | Article | - |
dc.identifier.doi | 10.1089/crispr.2022.0071 | - |
dc.identifier.bibliographicCitation | CRISPR JOURNAL, v.6, no.1, pp 52 - 61 | - |
dc.description.isOpenAccess | Y | - |
dc.identifier.wosid | 000904996500001 | - |
dc.identifier.scopusid | 2-s2.0-85148716403 | - |
dc.citation.endPage | 61 | - |
dc.citation.number | 1 | - |
dc.citation.startPage | 52 | - |
dc.citation.title | CRISPR JOURNAL | - |
dc.citation.volume | 6 | - |
dc.type.docType | Article | - |
dc.publisher.location | 미국 | - |
dc.subject.keywordPlus | CLEAVAGE | - |
dc.relation.journalResearchArea | Genetics & Heredity | - |
dc.relation.journalWebOfScienceCategory | Genetics & Heredity | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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