Article
Biochemistry & Molecular Biology
Zhongsen Li, Rui Ma, Dan Liu, Mingyue Wang, Ting Zhu, Yanxue Deng
Summary: CRISPR Cas9-mediated genome editing and base editing technologies have shown promising results in plant gene editing, with prime editing demonstrating high efficiency and precision. However, addressing the issue of inconsistent editing efficiencies across different target sites remains a challenge.
Article
Biotechnology & Applied Microbiology
Shengyao Zhi, Yuxi Chen, Guanglan Wu, Jinkun Wen, Jinni Wu, Qianyi Liu, Yang Li, Rui Kang, Sihui Hu, Jiahui Wang, Puping Liang, Junjiu Huang
Summary: Prime editor (PE) is a new genome editing tool that has the potential to correct the majority of known human genetic disease-related mutations. In this study, split-PEs were constructed and delivered using dual adenoassociated viruses (AAVs), successfully mediating gene editing in human cells and adult mouse retina.
Editorial Material
Plant Sciences
Niaz Ahmad, Muhammad Jawad Akbar Awan, Shahid Mansoor
Summary: Prime editing allows targeted changes in the genome without double-stranded breaks. However, its efficiency in plants is low. A recent study by Zong et al. improved prime-editing efficiency in both protoplasts and transgenic plants through a simple strategy.
TRENDS IN PLANT SCIENCE
(2023)
Article
Multidisciplinary Sciences
Xiangyang Li, Guiquan Zhang, Shisheng Huang, Yao Liu, Jin Tang, Mingtian Zhong, Xin Wang, Wenjun Sun, Yuan Yao, Quanjiang Ji, Xiaolong Wang, Jianghuai Liu, Shiqiang Zhu, Xingxu Huang
Summary: Strategies to improve the specificity of nuclease-based prime editor (PEn) are needed. Here the authors report a 53BP1-inhibitory ubiquitin variant-assisted PEn platform (uPEn) to inhibit NHEJ and enable precise prime editing for generation of insertions, deletions, and replacements.
NATURE COMMUNICATIONS
(2023)
Article
Biotechnology & Applied Microbiology
Yuan Zong, Yijing Liu, Chenxiao Xue, Boshu Li, Xiangyang Li, Yanpeng Wang, Ji Li, Guanwen Liu, Xingxu Huang, Xiaofeng Cao, Caixia Gao
Summary: In this study, we have improved the editing efficiency of prime editing by engineering the structure of the prime editor and combining it with specific guide RNAs. The optimized prime editor showed significantly increased editing efficiency in plant cells, and when combined with engineered guide RNAs, it resulted in the generation of herbicide-tolerant rice plants.
NATURE BIOTECHNOLOGY
(2022)
Article
Multidisciplinary Sciences
Qichen Yuan, Xue Gao
Summary: This study introduces a novel method for multiplex gene editing in human cells using CRISPR array architecture and RNA polymerase III promoter, enabling simultaneous editing of multiple disease-related genomic loci.
NATURE COMMUNICATIONS
(2022)
Review
Biochemical Research Methods
Md Mahmudul Hassan, Guoliang Yuan, Yang Liu, Mobashwer Alam, Carrie A. Eckert, Gerald A. Tuskan, John F. Golz, Xiaohan Yang
Summary: This article reviews the emerging strategies and existing technologies for improving plant genome editing. Several improvements have been proposed to address the inefficiency of gene targeting and prime editing in plants, such as RNA donor templating, chemically modified donor DNA template, and tandem-repeat homology-directed repair for gene targeting, and improved prime editing gRNA design, use of engineered reverse transcriptase enzymes, and splitting prime editing components for prime editing. These advancements are expected to drive the development of robust precision genome editing technologies for plants.
BIOTECHNOLOGY JOURNAL
(2022)
Review
Genetics & Heredity
Dawei Wang, Xiude Fan, Mengzhu Li, Tianbo Liu, Peng Lu, Guangxin Wang, Yuan Li, JunMing Han, JiaJun Zhao
Summary: The recently established prime editor (PE) system is considered as the next-generation gene-editing technology with higher flexibility and editing precision. This study introduces the basic principles of PE and reviews its recent progress in editing versatility, specificity, and efficiency in mammals. Furthermore, key considerations regarding the selection of PE variants, pegRNA design rules, and the evaluation of PE efficiency and accuracy are summarized.
Review
Biotechnology & Applied Microbiology
Gregory A. Newby, David R. Liu
Summary: Recent advances in genome editing technologies have made single-dose cures for genetic diseases more promising. Base editors and prime editors offer greater precision and efficiency compared to nuclease-dependent editing approaches, with less unwanted byproducts. Delivery of these editors in animal models has shown promise for in vivo therapeutic genome editing.
Article
Biochemistry & Molecular Biology
Peter J. Chen, Jeffrey A. Hussmann, Jun Yan, Friederike Knipping, Purnima Ravisankar, Pin-Fang Chen, Cidi Chen, James W. Nelson, Gregory A. Newby, Mustafa Sahin, Mark J. Osborn, Jonathan S. Weissman, Britt Adamson, David R. Liu
Summary: The study found that DNA mismatch repair (MMR) hinders prime editing and promotes undesired byproducts, but the efficiency of substitution, small insertion, and small deletion edits can be enhanced by developing PE4 and PE5 prime editing systems. Strategic silent mutations near the intended edit and optimization of prime editor protein also contribute to improved editing outcomes.
Article
Biochemistry & Molecular Biology
Yifu Tian, Dating Zhong, Xinbo Li, Rundong Shen, Han Han, Yuqin Dai, Qi Yao, Xuening Zhang, Qi Deng, Xuesong Cao, Jian-Kang Zhu, Yuming Lu
Summary: A geminivirus-mediated surrogate system called WDV-Gate was developed for high-throughput genome editing, which significantly increased the editing frequency, plantlet regeneration efficiency, and numbers of homozygous-edited plants compared to the conventional method. WDV-Gate was also demonstrated to be feasible for sequence knock-in for protein tagging.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Pei Ni, Yidi Zhao, Ximeng Zhou, Zehua Liu, Zhengwei Huang, Zhongfu Ni, Qixin Sun, Yuan Zong
Summary: Researchers developed an upgraded engineered plant prime editor, ePPEplus, by introducing a V223A substitution into reverse transcriptase in hexaploid wheat, which enhances editing efficiency by an average of 33.0-fold compared to the original PPE and 6.4-fold compared to ePPE. Importantly, they established a robust multiplex prime editing platform for simultaneous editing of multiple genes, achieving editing frequencies up to 74.5% in regenerated wheat plants, thus expanding the applicability of prime editors for stacking agronomic traits.
Article
Biochemistry & Molecular Biology
Yuan Zhuang, Jiangle Liu, Hao Wu, Qingguo Zhu, Yongchang Yan, Haowei Meng, Peng R. Chen, Chengqi Yi
Summary: The HOPE method utilizes paired pegRNAs encoding the same edits to achieve high editing efficiency in human embryonic kidney and colorectal carcinoma cells, showing improved product purity compared to the original PE3 system. This enhanced tool has the potential to broaden both fundamental research and therapeutic applications of prime editing.
NATURE CHEMICAL BIOLOGY
(2022)
Article
Multidisciplinary Sciences
Chunwei Zheng, Bin Liu, Xiaolong Dong, Nicholas Gaston, Erik J. Sontheimer, Wen Xue
Summary: Targeted insertion of large DNA fragments using prime editing (PE) has shown promise for genome engineering and gene therapy. However, the efficiency of inserting larger fragments (>400bp) remains low and in vivo application has not been demonstrated. Inspired by retrotransposons, researchers have developed a template-jumping (TJ) PE approach which allows for the insertion of large DNA fragments with high efficiency and without double-stranded DNA breaks.
NATURE COMMUNICATIONS
(2023)
Article
Biochemistry & Molecular Biology
Rui Tao, Yanhong Wang, Yun Hu, Yaoge Jiao, Lifang Zhou, Lurong Jiang, Li Li, Xingyu He, Min Li, Yamei Yu, Qiang Chen, Shaohua Yao
Summary: A novel prime editor, WT-PE, was designed in this study by fusing reverse transcriptase (RT) to nuclease wild-type Cas9, enabling efficient editing of large genomic fragments. The WT-PE system introduced a double strand break (DSB) and a single 3' extended flap in the target site, and coupled with paired prime editing guide RNAs (pegRNAs), it achieved bi-directional prime editing, allowing versatile and efficient large-scale genome editing. This WT-PE system has great potential for modeling or treating diseases related to large-fragment aberrations.
SIGNAL TRANSDUCTION AND TARGETED THERAPY
(2022)
Article
Plant Sciences
Shaoya Li, Jingying Li, Jiahui Zhang, Wenming Du, Jindong Fu, Suhas Sutar, Yunde Zhao, Lanqin Xia
JOURNAL OF EXPERIMENTAL BOTANY
(2018)
Letter
Biotechnology & Applied Microbiology
Yongwei Sun, Caroline Sparks, Huw Jones, Mandy Riley, Frederic Francis, Wenming Du, Lanqin Xia
PLANT BIOTECHNOLOGY JOURNAL
(2019)
Article
Biotechnology & Applied Microbiology
Shaoya Li, Jingying Li, Yubing He, Meilian Xu, Jiahui Zhang, Wenming Du, Yunde Zhao, Lanqin Xia
NATURE BIOTECHNOLOGY
(2019)
Letter
Biotechnology & Applied Microbiology
Shaoya Li, Yingxiao Zhang, Lanqin Xia, Yiping Qi
PLANT BIOTECHNOLOGY JOURNAL
(2020)
Letter
Biochemistry & Molecular Biology
Huiyuan Li, Jingying Li, Jilin Chen, Lei Yan, Lanqin Xia
Editorial Material
Biochemistry & Molecular Biology
Jingying Li, Huiyuan Li, Jilin Chen, Lei Yan, Lanqin Xia
Article
Biotechnology & Applied Microbiology
Tengfei Xia, Yanping Yang, Hongyuan Zheng, Xinyun Han, Huaibing Jin, Zijun Xiong, Weiqiang Qian, Lanqi Xia, Xiang Ji, Guangwei Li, Daowen Wang, Kunpu Zhang
Summary: LRK10-like receptor kinases (LRK10L-RLKs) are widely present in higher plants, but the understanding of their expression and function is still limited. Through transgenic wheat research, it was found that TtdLRK10L-1 is regulated in its expression by powdery mildew pathogens and exhibits resistance.
PLANT BIOTECHNOLOGY JOURNAL
(2021)
Article
Biotechnology & Applied Microbiology
Jingying Li, Guiai Jiao, Yongwei Sun, Jun Chen, Yingxin Zhong, Lei Yan, Dong Jiang, Youzhi Ma, Lanqin Xia
Summary: This study successfully generated high-amylose wheat through targeted mutagenesis of TaSBEIIa in modern wheat varieties using CRISPR/Cas9 technology, which increased starch and resistant starch content for health benefits.
PLANT BIOTECHNOLOGY JOURNAL
(2021)
Letter
Biochemistry & Molecular Biology
Jinman Luo, Shaoya Li, Jiajing Xu, Lei Yan, Youzhi Ma, Lanqin Xia
Article
Plant Sciences
Jilin Chen, Shaoya Li, Yubing He, Jingying Li, Lanqin Xia
Summary: This article summarizes the latest developments in CRISPR/Cas-mediated homology-directed repair technology for plants, highlighting significant achievements in crop improvement. It describes various strategies to enhance repair efficiency, as well as the challenges and open questions in precision genome editing for both plant biological research and crop improvement.
Review
Biochemistry & Molecular Biology
Jingying Li, Chen Zhang, Yubing He, Shaoya Li, Lei Yan, Yucai Li, Ziwei Zhu, Lanqin Xia
Summary: Precise replacement of an allele with an elite allele controlling an important agronomic trait in a predefined manner by gene editing technologies is highly desirable in crop improvement. Base editing and prime editing are two newly developed precision gene editing systems which can introduce the substitution of a single base and install the desired short indels to the target loci in the absence of double-strand breaks and donor repair templates, respectively. Various strategies have been attempted to optimize both base editor (BE) and prime editor (PE) to improve their editing efficacy, specificity, and expand the targeting scopes. Based on the latest development, appropriate BEs and PEs are recommended for basic plant research and crop improvement, and further optimization of these editors is proposed. Both BEs and PEs are envisioned to become routine and customized precise gene editing tools for plant biological research and crop improvement in the near future.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
