美国德克萨斯大学Ilya J. Finkelstein小组提出了用于精确基因组编辑的逆转录酶的发现和工程。这一研究成果于2025年10月23日发表在国际顶尖学术期刊《自然—生物技术》上。

在这里，该研究团队报告的发现和工程高效逆转录基因编辑的哺乳动物细胞和脊椎动物。通过元基因组数据的生物信息学分析和功能筛选，课题组确定了在哺乳动物细胞中高度活跃的逆转录逆转录酶。合理的设计进一步提高了编辑效率，达到了与传统的单链寡脱氧核苷酸供体相当的水平，但来自基因编码的磁带。逆转录编辑器表现出与Cas12a核酸酶和Cas9缺口酶的紧密活性，扩大了基因组靶标范围，并绕过了DNA双链断裂的需要。使用合理设计的逆转录编辑器，该研究组将分裂的GFP表位标签用于活细胞成像。最后，研究团队开发了一种全RNA递送策略，使细胞和脊椎动物胚胎中的无DNA基因编辑成为可能。这项工作建立了逆转录编辑器作为一种多功能和有效的精确基因编辑工具。

据介绍，逆转录酶可以通过自引反转录在细胞内产生多拷贝单链DNA。然而，它们在真核生物中插入遗传货物的潜力在很大程度上仍未被探索。

附：英文原文

Title: Discovery and engineering of retrons for precise genome editing

Author: Buffington, Jesse D., Kuo, Hung-Che, Hu, Kuang, Chang, You-Chiun, Javanmardi, Kamyab, Voigt, Brittney, Li, Yi-Ru, Little, Mary E., Devanathan, Sravan K., Xhemale, Blerta, Gray, Ryan S., Finkelstein, Ilya J.

Issue&Volume: 2025-10-23

Abstract: Retrons can produce multicopy single-stranded DNA in cells through self-primed reverse transcription. However, their potential for inserting genetic cargos in eukaryotes remains largely unexplored. Here we report the discovery and engineering of highly efficient retron-based gene editors for mammalian cells and vertebrates. Through bioinformatic analysis of metagenomic data and functional screening, we identify retron reverse transcriptases that are highly active in mammalian cells. Rational design further improves the editing efficiency to levels comparable with conventional single-stranded oligodeoxynucleotide donors but from a genetically encoded cassette. Retron editors exhibit robust activity with Cas12a nuclease and Cas9 nickase, expanding the genomic target scope and bypassing the need for a DNA double-stranded break. Using a rationally engineered retron editor, we incorporate a split GFP epitope tag for live-cell imaging. Lastly, we develop an all-RNA delivery strategy to enable DNA-free gene editing in cells and vertebrate embryos. This work establishes retron editors as a versatile and efficient tool for precise gene editing.

DOI: 10.1038/s41587-025-02879-3

Source: https://www.nature.com/articles/s41587-025-02879-3