Engineering self-deliverable ribonucleoproteins for genome editing in the brain.

Autor: Chen K; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA., Stahl EC; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA.; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA., Kang MH; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA.; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA., Xu B; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA., Allen R; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA., Trinidad M; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.; Innovative Genomics Institute, University of California, Berkeley, CA, USA.; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA., Doudna JA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.; Innovative Genomics Institute, University of California, Berkeley, CA, USA. doudna@berkeley.edu.; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.; Gladstone Institutes, San Francisco, CA, USA. doudna@berkeley.edu.; Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA. doudna@berkeley.edu.; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA. doudna@berkeley.edu.; Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Feb 26; Vol. 15 (1), pp. 1727. Date of Electronic Publication: 2024 Feb 26.
DOI: 10.1038/s41467-024-45998-2
Abstrakt: The delivery of CRISPR ribonucleoproteins (RNPs) for genome editing in vitro and in vivo has important advantages over other delivery methods, including reduced off-target and immunogenic effects. However, effective delivery of RNPs remains challenging in certain cell types due to low efficiency and cell toxicity. To address these issues, we engineer self-deliverable RNPs that can promote efficient cellular uptake and carry out robust genome editing without the need for helper materials or biomolecules. Screening of cell-penetrating peptides (CPPs) fused to CRISPR-Cas9 protein identifies potent constructs capable of efficient genome editing of neural progenitor cells. Further engineering of these fusion proteins establishes a C-terminal Cas9 fusion with three copies of A22p, a peptide derived from human semaphorin-3a, that exhibits substantially improved editing efficacy compared to other constructs. We find that self-deliverable Cas9 RNPs generate robust genome edits in clinically relevant genes when injected directly into the mouse striatum. Overall, self-deliverable Cas9 proteins provide a facile and effective platform for genome editing in vitro and in vivo.
(© 2024. The Author(s).)
Databáze: MEDLINE
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