Programming PAM antennae for efficient CRISPR-Cas9 DNA editing
文献类型：期刊论文
| 作者 | Wang, F; Hao, YY; Li, Q; Li, J; Zhang, HL; Zhang, XL; Wang, LH; Bustamante, C; Fan, CH |
| 刊名 | SCIENCE ADVANCES
 |
| 出版日期 | 2020 |
| 卷号 | 6期号:19页码:- |
| 关键词 | CAS9 |
| ISSN号 | 2375-2548 |
| DOI | 10.1126/sciadv.aay9948 |
| 文献子类 | 期刊论文 |
| 英文摘要 | Bacterial CRISPR-Cas9 nucleases have been repurposed as powerful genome editing tools. Whereas engineering guide RNAs or Cas nucleases have proven to improve the efficiency of CRISPR editing, modulation of protospaceradjacent motif (PAM), indispensable for CRISPR, has been less explored. Here, we develop a DNA origami-based platform to program a PAM antenna microenvironment and address its performance at the single-molecule level with submolecular resolution. To mimic spatially controlled in vivo PAM distribution as may occur in chromatin, we investigate the effect of PAM antennae surrounding target DNA. We find that PAM antennae effectively sensitize the DNA cleavage by recruiting Cas9 molecules. Super-resolution tracking of single single-guide RNA/Cas9s reveals localized translocation of Cas9 among spatially proximal PAMs. We find that the introduction of the PAM antennae effectively modulates the microenvironment for enhanced target cleavage (up to -50%). These results provide insight into factors that promote more efficient genome editing. |
| 语种 | 英语 |
| 源URL | [http://ir.sinap.ac.cn/handle/331007/33091]  |
| 专题 | 上海应用物理研究所_中科院上海应用物理研究所2011-2017年 |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Adv Res Inst, Zhangjiang Lab, Shanghai Synchrotron Radiat Facil, Shanghai 201210, Peoples R China 2.Southern Med Univ, Shanghai Jiao Tong Univ, Affiliated Peoples Hosp 6, Joint Res Ctr Precis Med,Affiliated Fengxian Hosp, South Campus, Shanghai 201499, Peoples R China 3.Shanghai Jiao Tong Univ, Frontiers Sci Ctr Transformat Mol, Sch Chem & Chem Engn, Shanghai 200240, Peoples R China 4.Shanghai Jiao Tong Univ, Inst Mol Med, Renji Hosp, Sch Med, Shanghai 200240, Peoples R China 5.Chinese Acad Sci, Shanghai Inst Appl Phys, CAS Key Lab Interfacial Phys & Technol, Shanghai 201800, Peoples R China 6.Univ Calif Berkeley, Howard Hughes Med Inst, Kavli Energy Nanosci Inst Berkeley, Dept Mol & Cell Biol,Dept Phys,Dept Chem,Inst Qua, Berkeley, CA 94720 USA 7.East China Normal Univ, Sch Chem & Mol Engn, Shanghai Key Lab Green Chem & Chem Proc, 500 Dongchuan Rd, Shanghai 200241, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, F,Hao, YY,Li, Q,et al. Programming PAM antennae for efficient CRISPR-Cas9 DNA editing[J]. SCIENCE ADVANCES,2020,6(19):-. |
| APA | Wang, F.,Hao, YY.,Li, Q.,Li, J.,Zhang, HL.,...&Fan, CH.(2020).Programming PAM antennae for efficient CRISPR-Cas9 DNA editing.SCIENCE ADVANCES,6(19),-. |
| MLA | Wang, F,et al."Programming PAM antennae for efficient CRISPR-Cas9 DNA editing".SCIENCE ADVANCES 6.19(2020):-. |
入库方式： OAI收割
来源：上海应用物理研究所
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