High-efficiency prime editing with optimized, paired pegRNAs in plants
Abstract
Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 °C leads to optimal performance in rice and that using two prime editing guide (peg) RNAs in trans encoding the same edits substantially enhances PE efficiency. Together, these approaches boost PE efficiency from 2.9-fold to 17.4-fold. Optimal pegRNAs or pegRNA pairs can be designed with our web application, PlantPegDesigner.
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CRISPR/Cas9 is a robust and versatile tool for precise mutation creation and trait improvement.
“In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a robust and versatile tool for precise mutation creation and trait improvement.”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Lin et al., 2021
More precise gene-editing technologies such as CRISPR/Cas9 gene editing, base editing, and prime editing are imperative to develop new barley lines with superior qualities and better performance under stress conditions.
“Therefore, more precise gene-editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) gene editing, base editing, and prime editing are imperative to develop new barley lines with superior qualities and better performance under stress conditions (Lin et al., 2021;Karunarathne et al., 2022).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Lin et al., 2021
Prime editing is the latest technique that can induce precise mutations.
“Prime editing is the latest technique that can induce precise mutations (Lin et al., 2021).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Lin et al., 2021
These single nucleotide polymorphisms (SNPs) can now be achieved through base or prime editing.
“These single nucleotide polymorphisms (SNPs) can now be achieved through base or prime editing (Lin et al., 2021;Xiong et al., 2022).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Lin et al., 2021
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References 22 · Cited by 325
- Authors
- Lin, Q., Jin, S., Zong, Y., Yu, H., Zhu, Z., Liu, G., Kou, L., Wang, Y., Qiu, J.-L., Li, J., & Gao, C.
- Affiliations
- Chinese Academy of Sciences (CN)
- Corresponding author
- Li, Jiayang
- Gao, Caixia
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2021-03-25
- Volume
- 39
- Issue
- 8
- Pages
- 923-927
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W3136535512
- Cited by
- 325
- References
- 22
- Citations by year
- 2026: 38 · 2025: 54 · 2024: 52 · 2023: 71 · 2022: 73 · 2021: 37
- FWCI
- 18.37
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
- Keywords
- prime editing, pegRNA, editing efficiency, melting point, rice, plant genome editing, web application, guide RNA design
- MeSH terms
- Gene Editing, RNA, Guide, CRISPR-Cas Systems, Oryza, RNA, Guide, Kinetoplastida, RNA, Plant, CRISPR-Cas Systems
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