Phage-assisted evolution of compact Cas9 variants targeting a simple NNG PAM
© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc..
Compact Cas9 nucleases hold great promise for therapeutic applications. Although several compact Cas9 nucleases have been developed, many genomic loci still could not be edited due to a lack of protospacer adjacent motifs (PAMs). We previously developed a compact SlugCas9 recognizing an NNGG PAM. Here we demonstrate that SlugCas9 displays comparable activity to SpCas9. We developed a simple phage-assisted evolution to engineer SlugCas9 for unique PAM requirements. Interestingly, we generated a SlugCas9 variant (SlugCas9-NNG) that could recognize an NNG PAM, expanding the targeting scope. We further developed a SlugCas9-NNG-based adenine base editor and demonstrated that it could be delivered by a single adeno-associated virus to disrupt PCSK9 splice donor and splice acceptor. These genome editors greatly enhance our ability for in vivo genome editing.
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:20 |
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| Enthalten in: |
Nature chemical biology - 20(2024), 3 vom: 06. März, Seite 344-352 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Qi, Tao [VerfasserIn] |
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| Links: |
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| Themen: |
Adenine |
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| Anmerkungen: |
Date Completed 04.03.2024 Date Revised 04.03.2024 published: Print-Electronic ErratumIn: Nat Chem Biol. 2024 Jan 5;:. - PMID 38182770 Citation Status MEDLINE |
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| doi: |
10.1038/s41589-023-01481-5 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM365438790 |
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| 520 | |a Compact Cas9 nucleases hold great promise for therapeutic applications. Although several compact Cas9 nucleases have been developed, many genomic loci still could not be edited due to a lack of protospacer adjacent motifs (PAMs). We previously developed a compact SlugCas9 recognizing an NNGG PAM. Here we demonstrate that SlugCas9 displays comparable activity to SpCas9. We developed a simple phage-assisted evolution to engineer SlugCas9 for unique PAM requirements. Interestingly, we generated a SlugCas9 variant (SlugCas9-NNG) that could recognize an NNG PAM, expanding the targeting scope. We further developed a SlugCas9-NNG-based adenine base editor and demonstrated that it could be delivered by a single adeno-associated virus to disrupt PCSK9 splice donor and splice acceptor. These genome editors greatly enhance our ability for in vivo genome editing | ||
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| 700 | 1 | |a Yang, Yuan |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Siqi |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Jingtong |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Qiang |e verfasserin |4 aut | |
| 700 | 1 | |a Tian, Yuwen |e verfasserin |4 aut | |
| 700 | 1 | |a Tang, Junnan |e verfasserin |4 aut | |
| 700 | 1 | |a Zheng, Wei V |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Yongming |e verfasserin |4 aut | |
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