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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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
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author 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
author_facet 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
author_sort Petri, Karl
collection PubMed
description 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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spelling pubmed-85538082022-02-16 CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells 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 Nat Biotechnol Article 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. 2022-02 2021-04-29 /pmc/articles/PMC8553808/ /pubmed/33927418 http://dx.doi.org/10.1038/s41587-021-00901-y Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
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
CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title_full CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title_fullStr CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title_full_unstemmed CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title_short CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
title_sort crispr prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
topic Article
url 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
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