Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells
Abstract Prime editing (PE) allows for precise genome editing in human pluripotent stem cells (hPSCs), such as introducing single nucleotide modifications, small insertions or deletions at a specific genomic locus. Here, we systematically compare a panel of prime editing conditions in hPSCs and gene...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-12-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55104-1 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841559306749607936 |
|---|---|
| author | Youjun Wu Aaron Zhong Mega Sidharta Tae Wan Kim Bernny Ramirez Benjamin Persily Lorenz Studer Ting Zhou |
| author_facet | Youjun Wu Aaron Zhong Mega Sidharta Tae Wan Kim Bernny Ramirez Benjamin Persily Lorenz Studer Ting Zhou |
| author_sort | Youjun Wu |
| collection | DOAJ |
| description | Abstract Prime editing (PE) allows for precise genome editing in human pluripotent stem cells (hPSCs), such as introducing single nucleotide modifications, small insertions or deletions at a specific genomic locus. Here, we systematically compare a panel of prime editing conditions in hPSCs and generate a potent prime editor, “PE-Plus”, through co-inhibition of mismatch repair and p53-mediated cellular stress responses. We further establish an inducible prime editing platform in hPSCs by incorporating the PE-Plus into a safe-harbor locus and demonstrated temporal control of precise editing in both hPSCs and differentiated cells. By evaluating disease-associated mutations, we show that this platform allows efficient creation of both monoallelic and biallelic disease-relevant mutations in hPSCs. In addition, this platform enables the efficient introduction of single or multiple edits in one step, demonstrating potential for multiplex editing. Our method presents an efficient and controllable multiplex prime editing tool in hPSCs and their differentiated progeny. |
| format | Article |
| id | doaj-art-aa11b2a3309b46949872cd9e9f48977d |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-aa11b2a3309b46949872cd9e9f48977d2025-01-05T12:35:36ZengNature PortfolioNature Communications2041-17232024-12-0115111310.1038/s41467-024-55104-1Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cellsYoujun Wu0Aaron Zhong1Mega Sidharta2Tae Wan Kim3Bernny Ramirez4Benjamin Persily5Lorenz Studer6Ting Zhou7The SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueThe SKI Stem Cell Research Facility, The Center for Stem Cell Biology and Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, 1275 York AvenueAbstract Prime editing (PE) allows for precise genome editing in human pluripotent stem cells (hPSCs), such as introducing single nucleotide modifications, small insertions or deletions at a specific genomic locus. Here, we systematically compare a panel of prime editing conditions in hPSCs and generate a potent prime editor, “PE-Plus”, through co-inhibition of mismatch repair and p53-mediated cellular stress responses. We further establish an inducible prime editing platform in hPSCs by incorporating the PE-Plus into a safe-harbor locus and demonstrated temporal control of precise editing in both hPSCs and differentiated cells. By evaluating disease-associated mutations, we show that this platform allows efficient creation of both monoallelic and biallelic disease-relevant mutations in hPSCs. In addition, this platform enables the efficient introduction of single or multiple edits in one step, demonstrating potential for multiplex editing. Our method presents an efficient and controllable multiplex prime editing tool in hPSCs and their differentiated progeny.https://doi.org/10.1038/s41467-024-55104-1 |
| spellingShingle | Youjun Wu Aaron Zhong Mega Sidharta Tae Wan Kim Bernny Ramirez Benjamin Persily Lorenz Studer Ting Zhou Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells Nature Communications |
| title | Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells |
| title_full | Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells |
| title_fullStr | Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells |
| title_full_unstemmed | Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells |
| title_short | Robust and inducible genome editing via an all-in-one prime editor in human pluripotent stem cells |
| title_sort | robust and inducible genome editing via an all in one prime editor in human pluripotent stem cells |
| url | https://doi.org/10.1038/s41467-024-55104-1 |
| work_keys_str_mv | AT youjunwu robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT aaronzhong robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT megasidharta robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT taewankim robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT bernnyramirez robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT benjaminpersily robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT lorenzstuder robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells AT tingzhou robustandinduciblegenomeeditingviaanallinoneprimeeditorinhumanpluripotentstemcells |