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journal article·2026-06-26Open access

Discovery and Engineering of a Rat Endogenous Retrovirus Reverse Transcriptase for Efficient Prime Editing

Linsha Ma, Pengcheng Yao, Shengqi Wu, Yuanyuan Shi, Lang Qin, Baitao Li, Jiayi Zhu, Minhua Huang, Yichong Zhu, Yuwen Song, Jinhuan Pang, Ziping Guo, Guochuan Wu, Chen Wang, Kewei Xu, Ruihua Huang, Quan Kuang, Liang Qu, Changtian Pan, Xianrong Xie, Qinlong Zhu, Jiaying Huang, Qiupeng Lin

Advanced Science
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Source abstract

Abstract

ABSTRACT CRISPR‐based prime editors (PEs) install precise edits into genomic DNA without generating double‐strand breaks. Their editing efficiency is highly dependent on reverse transcriptases (RTs), but efficient RT candidates remain limited. Here, we identified 19 novel active RTs by screening 558 candidates. Among them, RERV‐RT, derived from Rattus norvegicus , exhibited the highest activity. Through structure‐guided engineering and deep mutational scanning, we developed an optimized variant, enRERV‐RT, which outperforms conventional M‐MLV‐RT‐based PE systems by 1.20‐fold in mammalian and plant cells, and by 1.88‐fold at hard‐to‐edit loci, while enabling precise multiplex editing of functionally relevant genes. Additionally, we developed a high‐throughput platform, TRAP‐seq‐PE, to systematically evaluate prime editor performance. Across diverse mutation types, we found that PE systems based on enRERV‐RT exhibited higher editing efficiencies than those based on M‐MLV‐RT. Collectively, our work establishes a versatile, high‐efficiency PE system, thereby facilitating advances in clinical gene therapy and precise crop breeding.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: CRISPR and Genetic Engineering · Pluripotent Stem Cells Research · RNA regulation and disease

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Guangdong Basic Research Center of Excellence for Precise Breeding of Future Crops Key Laboratory for Enhancing Resource Use Efficiency of Crops in South China State Key Laboratory For Conservation and Utilization of Subtropical Agro‐Bioresources Guangdong Provincial Key Laboratory of Plant Molecular Breeding Ministry of Agriculture and Rural Affairs College of Agriculture South China Agricultural University Guangzhou China

Department of Radiology The Affiliated Panyu Central Hospital GMU‐GIBH Joint School of Life Sciences Guangdong Provincial Key Laboratory of Protein Modification and Disease The Guangdong‐Hong Kong‐Macao Joint Laboratory for Cell Fate Regulation and Diseases Guangzhou Medical University Guangzhou China

State Key Laboratory of Respiratory Disease National Clinical Research Center for Respiratory Disease National Center for Respiratory Medicine Joint International Research Laboratory of Respiratory Health Guangdong Basic Research Center of Excellence for Respiratory Medicine Guangzhou Institute of Respiratory Health Guangzhou Medical University Guangzhou China

College of Fisheries Southwest University Chongqing China

Department of Life Sciences Beijing Normal‐Hong Kong Baptist University Zhuhai China

Dermatology Hospital of Southern Medical University Guangzhou China

Guangdong Key Laboratory of Biotechnology for Plant Development College of Life Sciences South China Normal University Guangzhou China

State Key Laboratory of Quantitative Synthetic Biology Shenzhen Institute of Synthetic Biology Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences Shenzhen China

College of Life Sciences Nanchang Normal University Nanchang China

Frontier Innovation Center Department of Systems Biology For Medicine Qidong‐Fudan Innovative Institution of Medical Sciences School of Basic Medical Sciences Fudan University Shanghai China

Clinical Center for Biotherapy Zhongshan Hospital Fudan University Shanghai China

Department of Horticulture College of Agriculture and Biotechnology Zhejiang University Hangzhou China

Key Laboratory of Horticultural Plant Growth Development and Quality Improvement Ministry of Agricultural Zhejiang University Hangzhou China

Institute of BioFoundry College of Life Sciences South China Agricultural University Guangzhou China

Funding

National Natural Science Foundation of China

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