論文 - 詳細
| RRC ID | 82147 |
|---|---|
| 著者 | Degtev D, Bravo J, Emmanouilidi A, Zdravković A, Choong OK, Liz Touza J, Selfjord N, Weisheit I, Francescatto M, Akcakaya P, Porritt M, Maresca M, Taylor D, Sienski G. |
| タイトル | Engineered PsCas9 enables therapeutic genome editing in mouse liver with lipid nanoparticles. |
| ジャーナル | Nat Commun |
| Abstract |
Clinical implementation of therapeutic genome editing relies on efficient in vivo delivery and the safety of CRISPR-Cas tools. Previously, we identified PsCas9 as a Type II-B family enzyme capable of editing mouse liver genome upon adenoviral delivery without detectable off-targets and reduced chromosomal translocations. Yet, its efficacy remains insufficient with non-viral delivery, a common challenge for many Cas9 orthologues. Here, we sought to redesign PsCas9 for in vivo editing using lipid nanoparticles. We solve the PsCas9 ribonucleoprotein structure with cryo-EM and characterize it biochemically, providing a basis for its rational engineering. Screening over numerous guide RNA and protein variants lead us to develop engineered PsCas9 (ePsCas9) with up to 20-fold increased activity across various targets and preserved safety advantages. We apply the same design principles to boost the activity of FnCas9, an enzyme phylogenetically relevant to PsCas9. Remarkably, a single administration of mRNA encoding ePsCas9 and its guide formulated with lipid nanoparticles results in high levels of editing in the Pcsk9 gene in mouse liver, a clinically relevant target for hypercholesterolemia treatment. Collectively, our findings introduce ePsCas9 as a highly efficient, and precise tool for therapeutic genome editing, in addition to the engineering strategy applicable to other Cas9 orthologues. |
| 巻・号 | 15(1) |
| ページ | 9173 |
| 公開日 | 2024-11-7 |
| DOI | 10.1038/s41467-024-53418-8 |
| PII | 10.1038/s41467-024-53418-8 |
| PMID | 39511150 |
| PMC | PMC11544209 |
| MeSH | Animals CRISPR-Associated Protein 9* / genetics CRISPR-Associated Protein 9* / metabolism CRISPR-Cas Systems* Cryoelectron Microscopy Gene Editing* / methods Genetic Therapy / methods HEK293 Cells Humans Lipids / chemistry Liposomes Liver* / metabolism Mice Mice, Inbred C57BL Nanoparticles* / chemistry Proprotein Convertase 9* / genetics Proprotein Convertase 9* / metabolism RNA, Guide, CRISPR-Cas Systems* / genetics |
| IF | 12.121 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 19 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HuH-7(RCB1366) |