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Small-molecule compounds boost genome-editing efficiency of cytosine base editor
Nucleic Acids Research ( IF 16.6 ) Pub Date : 2021-07-17 , DOI: 10.1093/nar/gkab645
Tianyuan Zhao 1 , Qing Li 2 , Chenchen Zhou 1 , Xiujuan Lv 1 , Hongyan Liu 1 , Tianxiang Tu 1 , Na Tang 3 , Yanbo Cheng 2 , Xiaoyu Liu 1 , Changbao Liu 4 , Junzhao Zhao 4 , Zongming Song 5 , Haoyi Wang 3 , Jinsong Li 2 , Feng Gu 1
Affiliation  

Cytosine base editor (CBE) enables targeted C-to-T conversions at single base-pair resolution and thus has potential therapeutic applications in humans. However, the low efficiency of the system limits practical use of this approach. We reported a high-throughput human cells-based reporter system that can be harnessed for quickly measuring editing activity of CBE. Screening of 1813 small-molecule compounds resulted in the identification of Ricolinostat (an HDAC6 inhibitor) that can enhance the efficiency of BE3 in human cells (2.45- to 9.21-fold improvement). Nexturastat A, another HDAC6 inhibitor, could also increase BE3-mediated gene editing by 2.18- to 9.95-fold. Ricolinostat and Nexturastat A also boost base editing activity of the other CBE variants (BE4max, YE1-BE4max, evoAPOBEC1-BE4max and SpRY-CBE4max, up to 8.32-fold). Meanwhile, combined application of BE3 and Ricolinostat led to >3-fold higher efficiency of correcting a pathogenic mutation in ABCA4 gene related to Stargardt disease in human cells. Moreover, we demonstrated that our strategy could be applied for efficient generation of mouse models through direct zygote injection and base editing in primary human T cells. Our study provides a new strategy to improve the activity and specificity of CBE in human cells. Ricolinostat and Nexturastat A augment the effectiveness and applicability of CBE.

中文翻译:

小分子化合物提高了胞嘧啶碱基编辑器的基因组编辑效率

胞嘧啶碱基编辑器 (CBE) 能够以单碱基对分辨率实现靶向 C 到 T 的转换,因此在人类中具有潜在的治疗应用。然而,系统的低效率限制了这种方法的实际使用。我们报告了一种基于人类细胞的高通量报告系统,可用于快速测量 CBE 的编辑活动。通过对 1813 种小分子化合物的筛选,鉴定出 Ricolinostat(一种 HDAC6 抑制剂)可以提高 BE3 在人体细胞中的效率(提高 2.45 至 9.21 倍)。另一种 HDAC6 抑制剂 Nexturastat A 也可以将 BE3 介导的基因编辑增加 2.18 至 9.95 倍。Ricolinostat 和 Nexturastat A 还提高了其他 CBE 变体(BE4max、YE1-BE4max、evoAPOBEC1-BE4max 和 SpRY-CBE4max,高达 8.32 倍)的碱基编辑活性。同时,BE3和Ricolinostat的联合应用导致纠正人类细胞中与Stargardt病相关的ABCA4基因的致病突变的效率提高了3倍以上。此外,我们证明了我们的策略可以通过直接受精卵注射和原代人类 T 细胞中的碱基编辑来有效生成小鼠模型。我们的研究提供了一种新的策略来提高 CBE 在人体细胞中的活性和特异性。Ricolinostat 和 Nexturastat A 增强了 CBE 的有效性和适用性。我们证明了我们的策略可以通过直接合子注射和原代人类 T 细胞中的碱基编辑来有效生成小鼠模型。我们的研究提供了一种新的策略来提高 CBE 在人体细胞中的活性和特异性。Ricolinostat 和 Nexturastat A 增强了 CBE 的有效性和适用性。我们证明了我们的策略可以通过直接合子注射和原代人类 T 细胞中的碱基编辑来有效生成小鼠模型。我们的研究提供了一种新的策略来提高 CBE 在人体细胞中的活性和特异性。Ricolinostat 和 Nexturastat A 增强了 CBE 的有效性和适用性。
更新日期:2021-07-17
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