دورية أكاديمية
Chimeric CRISPR-CasX enzymes and guide RNAs for improved genome editing activity.
| العنوان: | Chimeric CRISPR-CasX enzymes and guide RNAs for improved genome editing activity. |
|---|---|
| المؤلفون: | Tsuchida CA; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, University of California, Berkeley, California 94720, USA; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA., Zhang S; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., Doost MS; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA., Zhao Y; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., Wang J; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., O'Brien E; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA., Fang H; Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China., Li CP; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., Li D; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., Hai ZY; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China., Chuck J; Department of Integrative Structural and Computational Biology, The Scripps Research Institute, Jupiter, Florida 33458, USA., Brötzmann J; Department of Structural Cell Biology, Max Planck Institute of Biochemistry, Martinsried, Germany., Vartoumian A; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA., Burstein D; School of Molecular Cell Biology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 6997801, Israel., Chen XW; Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China., Nogales E; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA; Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA; Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA; Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA., Doudna JA; University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, University of California, Berkeley, California 94720, USA; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA; Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA; Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA; Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA; Department of Chemistry, University of California, Berkeley, California 94720, USA; Gladstone Institute of Data Science and Biotechnology. Gladstone Institutes, San Francisco, California 94158, USA. Electronic address: doudna@berkeley.edu., Liu JG; Beijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China. Electronic address: junjiegogoliu@tsinghua.edu.cn. |
| المصدر: | Molecular cell [Mol Cell] 2022 Mar 17; Vol. 82 (6), pp. 1199-1209.e6. Date of Electronic Publication: 2022 Feb 25. |
| نوع المنشور: | Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 9802571 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1097-4164 (Electronic) Linking ISSN: 10972765 NLM ISO Abbreviation: Mol Cell Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Cambridge Ma : Cell Press Original Publication: Cambridge, Mass. : Cell Press, c1997- |
| مواضيع طبية MeSH: | Gene Editing*/methods , RNA, Guide, CRISPR-Cas Systems*/genetics , RNA, Guide, CRISPR-Cas Systems*/metabolism, Animals ; CRISPR-Cas Systems/genetics ; Endonucleases/genetics ; Endonucleases/metabolism ; Mammals/metabolism ; RNA/genetics |
| مستخلص: | A compact protein with a size of <1,000 amino acids, the CRISPR-associated protein CasX is a fundamentally distinct RNA-guided nuclease when compared to Cas9 and Cas12a. Although it can induce RNA-guided genome editing in mammalian cells, the activity of CasX is less robust than that of the widely used S. pyogenes Cas9. Here, we show that structural features of two CasX homologs and their guide RNAs affect the R-loop complex assembly and DNA cleavage activity. Cryo-EM-based structural engineering of either the CasX protein or the guide RNA produced two new CasX genome editors (DpbCasX-R3-v2 and PlmCasX-R1-v2) with significantly improved DNA manipulation efficacy. These results advance both the mechanistic understanding of CasX and its application as a genome-editing tool. Competing Interests: Declaration of interests J.A.D., E.N., J.J.G.L., C.A.T., M.S.D., and E.O. have filed a related patent on CasX mutations and new guide RNAs described herein with the United States Patent and Trademark Office. J.A.D. is a co-founder of Caribou Biosciences, Editas Medicine, Intellia Therapeutics, Scribe Therapeutics, and Mammoth Biosciences, and a director of Johnson & Johnson. J.A.D is a scientific advisor to Caribou Biosciences, Intellia Therapeutics, eFFECTOR Therapeutics, Scribe Therapeutics, Synthego, and Inari. (Copyright © 2022 Elsevier Inc. All rights reserved.) |
| التعليقات: | Comment in: Mol Cell. 2022 Mar 17;82(6):1083-1085. (PMID: 35303481) |
| References: | Elife. 2016 Nov 15;5:. (PMID: 27845625) Cell. 2016 Dec 15;167(7):1814-1828.e12. (PMID: 27984729) Nature. 2015 Apr 9;520(7546):186-91. (PMID: 25830891) Nat Methods. 2017 Mar;14(3):290-296. (PMID: 28165473) Cell. 2018 Mar 8;172(6):1239-1259. (PMID: 29522745) Annu Rev Biophys. 2017 May 22;46:505-529. (PMID: 28375731) Nucleic Acids Res. 2021 Apr 19;49(7):4120-4128. (PMID: 33764415) Acta Crystallogr D Struct Biol. 2019 Oct 1;75(Pt 10):861-877. (PMID: 31588918) Clin Transl Med. 2021 Sep;11(9):e537. (PMID: 34586743) Nat Med. 2019 Feb;25(2):249-254. (PMID: 30692695) Mol Cell. 2021 Oct 21;81(20):4333-4345.e4. (PMID: 34480847) Cell. 2020 Apr 2;181(1):136-150. (PMID: 32243786) Cell. 2016 Jan 14;164(1-2):29-44. (PMID: 26771484) FEBS J. 2016 Sep;283(17):3162-9. (PMID: 27234458) Nat Biotechnol. 2019 Mar;37(3):276-282. (PMID: 30742127) Nucleic Acids Res. 2016 Jul 8;44(W1):W351-5. (PMID: 27131377) Methods. 2009 Oct;49(2):174-80. (PMID: 19398010) Science. 2018 Apr 27;360(6387):436-439. (PMID: 29449511) Cell. 2015 Oct 22;163(3):759-71. (PMID: 26422227) Nat Commun. 2018 Aug 29;9(1):3497. (PMID: 30158648) ACS Synth Biol. 2019 Oct 18;8(10):2228-2237. (PMID: 31532637) RNA Biol. 2019 Apr;16(4):380-389. (PMID: 30856357) Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21. (PMID: 20057044) Mol Cell. 2021 Feb 4;81(3):558-570.e3. (PMID: 33333018) Annu Rev Virol. 2014 Nov;1(1):427-51. (PMID: 26958729) Curr Opin Microbiol. 2017 Jun;37:67-78. (PMID: 28605718) Nature. 2020 Feb;578(7794):229-236. (PMID: 32051598) Cell Res. 2019 May;29(5):345-346. (PMID: 30992542) BMC Genomics. 2019 Jun 27;20(1):528. (PMID: 31248380) Nat Rev Drug Discov. 2021 Mar;20(3):173-174. (PMID: 33495615) Nat Chem Biol. 2021 Nov;17(11):1132-1138. (PMID: 34475565) Science. 2020 Jul 17;369(6501):333-337. (PMID: 32675376) Nature. 2017 Feb 9;542(7640):237-241. (PMID: 28005056) Cell. 2016 May 5;165(4):949-62. (PMID: 27114038) Nature. 2019 Feb;566(7743):218-223. (PMID: 30718774) Nat Rev Microbiol. 2020 Feb;18(2):67-83. (PMID: 31857715) Nat Biotechnol. 2018 Apr;36(4):324-327. (PMID: 29553573) Protein Sci. 2020 Apr;29(4):1069-1078. (PMID: 31730249) Nat Biotechnol. 2022 Jan;40(1):94-102. (PMID: 34475560) Nucleic Acids Res. 2020 May 21;48(9):5037-5053. (PMID: 32315032) Science. 2014 Nov 28;346(6213):1258096. (PMID: 25430774) Nat Methods. 2017 Apr;14(4):331-332. (PMID: 28250466) |
| معلومات مُعتمدة: | F31 HL156468 United States HL NHLBI NIH HHS; P01 GM051487 United States GM NIGMS NIH HHS; United States HHMI Howard Hughes Medical Institute |
| فهرسة مساهمة: | Keywords: CRISPR; Cas12e; CasX; DNA cleavage; RNA-guided DNA nuclease; cryo-EM; genome editing; nucleic acid manipulation; sgRNA; structural engineering |
| المشرفين على المادة: | 0 (RNA, Guide, CRISPR-Cas Systems) 63231-63-0 (RNA) EC 3.1.- (Endonucleases) |
| تواريخ الأحداث: | Date Created: 20220227 Date Completed: 20220413 Latest Revision: 20240104 |
| رمز التحديث: | 20240104 |
| مُعرف محوري في PubMed: | PMC9189900 |
| DOI: | 10.1016/j.molcel.2022.02.002 |
| PMID: | 35219382 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1097-4164 |
|---|---|
| DOI: | 10.1016/j.molcel.2022.02.002 |