Extended pegRNAs enhance the editing capability of Prime editing.

IF 14.3 1区工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Trends in biotechnology Pub Date : 2024-09-27 DOI:10.1016/j.tibtech.2024.09.004

Kezhang He, Qiaomei Xue, Wei Zhou, Pengqi Wang, Xiaodan Hu, Tongtong Lin, Nan Chen, Bowen Wang, Tianhua Ma, Sheng Ding

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引用次数: 0

Abstract

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.

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扩展的 pegRNA 增强了 Prime 编辑的编辑能力。

基因组编辑在生物医学研究中极具价值。尽管基因组编辑（PE）技术用途广泛，但目前的基因组编辑（PE）技术仅限于短序列改变[最多约 44 碱基对 (bp)]，而且在整个基因组位点上表现出不一致或低效率，尤其是在面对多 T 序列时。为了应对这些挑战，我们开发了一种扩展 PE（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PE引导RNA（exegRNA），exPE大大提高了编辑效率，克服了多T序列带来的挑战。与传统 PE 相比，exPE 将碱基转换和短插入的效率提高了 14 倍，在多 T 序列区域的效率更是显著提高了 259 倍。与众不同的是，exPE 能够将基因大小的 DNA 片段无缝插入基因组，有可能纠正近 90% 的人类基因变异，从而拓宽了其在基因研究和治疗中的应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Trends in biotechnology 工程技术-生物工程与应用微生物

CiteScore

28.60

自引率

1.20%

发文量

198

审稿时长

1 months

期刊介绍： Trends in Biotechnology publishes reviews and perspectives on the applied biological sciences, focusing on useful science applied to, derived from, or inspired by living systems. The major themes that TIBTECH is interested in include: Bioprocessing (biochemical engineering, applied enzymology, industrial biotechnology, biofuels, metabolic engineering) Omics (genome editing, single-cell technologies, bioinformatics, synthetic biology) Materials and devices (bionanotechnology, biomaterials, diagnostics/imaging/detection, soft robotics, biosensors/bioelectronics) Therapeutics (biofabrication, stem cells, tissue engineering and regenerative medicine, antibodies and other protein drugs, drug delivery) Agroenvironment (environmental engineering, bioremediation, genetically modified crops, sustainable development).