CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells

Prime Editors have been delivered using DNA or mRNA vectors. Here we demonstrate prime editing (PE) with purified ribonucleoprotein (RNP) complexes. We introduced somatic mutations in zebrafish embryos with frequencies as high as 30% and demonstrate germline transmission. We also observed unintended...

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Detalles Bibliográficos
Autores principales: Petri, Karl, Zhang, Weiting, Ma, Junyan, Schmidts, Andrea, Lee, Hyunho, Horng, Joy E., Kim, Daniel Y., Kurt, Ibrahim C., Clement, Kendell, Hsu, Jonathan Y., Pinello, Luca, Maus, Marcela V., Joung, J. Keith, Yeh, Jing-Ruey Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2022
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8553808/
https://www.ncbi.nlm.nih.gov/pubmed/33927418
http://dx.doi.org/10.1038/s41587-021-00901-y
Descripción
Sumario:Prime Editors have been delivered using DNA or mRNA vectors. Here we demonstrate prime editing (PE) with purified ribonucleoprotein (RNP) complexes. We introduced somatic mutations in zebrafish embryos with frequencies as high as 30% and demonstrate germline transmission. We also observed unintended insertions, deletions and pegRNA scaffold incorporations. In HEK293T and primary human T cells, PE RNP complexes introduced desired edits with frequencies of up to 21% and 7.5%, respectively.

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