지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제30권 제12호
- 발행연도
- 2020.1
- 수록면
- 1,919 - 1,926 (8page)
이용수
초록· 키워드
오류제보하기
CRISPR interference (CRISPRi) has been developed as a transcriptional control tool by inactivating the DNA cleavage ability of Cas9 nucleases to produce dCas9 (deactivated Cas9), and leaving dCas9 the ability to specifically bind to the target DNA sequence. CRISPR/Cas9 technology has limitations in designing target-specific single-guide RNA (sgRNA) due to the dependence of protospacer adjacent motif (PAM) (5'-NGG) for binding target DNAs. Reportedly, Cas9-NG recognizing 5'-NG as the PAM sequence has been constructed by removing the dependence on the last base G of PAM through protein engineering of Cas9. In this study, a dCas9-NG protein was engineered by introducing two active site mutations in Cas9-NG, and its ability to regulate transcription was evaluated in the gal promoter in E. coli. Analysis of cell growth rate, D-galactose consumption rate, and gal transcripts confirmed that dCas9-NG can completely repress the promoter by recognizing DNA targets with PAM of 5'-NGG, NGA, NGC, NGT, and NAG. Our study showed possible PAM sequences for dCas9-NG and provided information on target-specific sgRNA design for regulation of both gene expression and cellular metabolism.
목차
등록된 정보가 없습니다.
참고문헌 (28)
참고문헌 신청
Horvath P / 2010 / CRISPR/Cas, the immune system of bacteria and archaea / Science 327:167~170
Barrangou R / 2007 / CRISPR provides acquired resistance against viruses in prokaryotes / Science 315:1709~1712
Terns MP / 2011 / CRISPR-based adaptive immune systems / Curr. Opin. Microbiol 14:321~327
Cox DBT / 2017 / RNA editing with CRISPR-Cas13 / Science 358:1019~1027
Anders C / 2014 / Structural basis of PAM-dependent target DNA recognition by the Cas9endonuclease / Nature 513:569~573
Barrangou R / 2007 / CRISPR provides acquired resistance against viruses in prokaryotes / Science 315:1709~1712
Terns MP / 2011 / CRISPR-based adaptive immune systems / Curr. Opin. Microbiol 14:321~327
Cox DBT / 2017 / RNA editing with CRISPR-Cas13 / Science 358:1019~1027
Anders C / 2014 / Structural basis of PAM-dependent target DNA recognition by the Cas9endonuclease / Nature 513:569~573
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