He Kezhang, Xue Qiaomei, Zhou Wei, Wang Pengqi, Hu Xiaodan, Lin Tongtong, Chen Nan, Wang Bowen, Ma Tianhua, Ding Sheng
New Cornerstone Science Laboratory, School of Pharmaceutical Sciences, Tsinghua University, Beijing 100084, China.
New Cornerstone Science Laboratory, School of Pharmaceutical Sciences, Tsinghua University, Beijing 100084, China.
Trends Biotechnol. 2025 Jan;43(1):206-219. doi: 10.1016/j.tibtech.2024.09.004. Epub 2024 Sep 27.
Genome editing is highly valuable in biomedical research. Despite their versatility, current Prime editing (PE) techniques are limited to short sequence alterations [up to ~44 base pairs (bp)], and exhibit inconsistent or low efficiency across genomic loci, particularly when faced with poly-T sequences. To address these challenges, we developed an extended PE (exPE) technique that can potentially execute any precise genome editing. By harnessing RNA polymerase II (Pol II) promoters to transcribe extended PE guide RNAs (expegRNAs), exPE substantially improves editing efficiency and overcomes the challenges posed by poly-T sequences. Compared with conventional PE, exPE achieves up to a 14-fold increase in the efficiency of base conversions and short insertions, and, remarkably, up to a 259-fold improvement in regions with poly-T sequences. Uniquely, exPE enables seamless insertion of gene-sized DNA fragments into genomes, potentially correcting nearly 90% of human genetic variants, thereby broadening its applications in genetic research and therapy.
基因组编辑在生物医学研究中具有极高的价值。尽管当前的先导编辑(PE)技术具有多功能性,但它们仅限于短序列改变[最多约44个碱基对(bp)],并且在整个基因组位点上表现出不一致或低效的情况,尤其是在面对多聚T序列时。为应对这些挑战,我们开发了一种扩展先导编辑(exPE)技术,它有可能执行任何精确的基因组编辑。通过利用RNA聚合酶II(Pol II)启动子转录扩展的PE引导RNA(expegRNAs),exPE显著提高了编辑效率,并克服了多聚T序列带来的挑战。与传统的PE相比,exPE在碱基转换和短插入效率上提高了多达14倍,而且在具有多聚T序列的区域中显著提高了多达259倍。独特的是,exPE能够将基因大小的DNA片段无缝插入基因组,有可能校正近90%的人类遗传变异,从而拓宽了其在基因研究和治疗中的应用。
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