Methods and Applications of CRISPR-Mediated Base Editing in Eukaryotic Genomes.

Methods and Applications of CRISPR-Mediated Base Editing in Eukaryotic Genomes.
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DOI:
10.1016/j.molcel.2017.09.029
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发表时间:
2017-10-05
期刊:
影响因子:
16
通讯作者:
Bassik MC
Bassik MC
中科院分区:
生物学1区
文献类型:
--
作者:
Hess GT;Tycko J;Yao D;Bassik MC

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在过去的几年里,CRISPR-Cas9系统的应用开发出现了爆炸式增长,从高效的基因组编辑到高通量筛选,再到一系列DNA和染色质修饰酶的招募。虽然与Cas9核酸酶切割偶联的同源定向修复(HDR)已被成功地用于修复和重写基因组,但最近开发的碱基编辑系统提供了一种有用的正交策略来工程化核苷酸取代。碱基编辑依赖于胞苷脱氨酶的募集来引入变化(而不是双链断裂和供体模板),并且提供了效率的潜在改进,同时限制了损伤并简化了编辑机器的递送。同时,这些系统使得新的诱变策略能够引入用于工程和发现的序列多样性。在这里,我们回顾了不同的碱基编辑平台,包括它们的脱氨酶招募策略和编辑结果,并将它们与其他CRISPR基因组编辑技术进行比较。此外,我们还讨论了这些系统如何应用于治疗,工程和研究环境。最后,我们探讨了这项新兴技术的未来发展方向。
The past several years have seen an explosion in development of applications for the CRISPR-Cas9 system, from efficient genome editing, to high-throughput screening, to recruitment of a range of DNA and chromatin-modifying enzymes. While homology-directed repair (HDR) coupled with Cas9 nuclease cleavage has been used with great success to repair and re-write genomes, recently developed base editing systems present a useful orthogonal strategy to engineer nucleotide substitutions. Base editing relies on recruitment of cytidine deaminases to introduce changes (rather than double stranded breaks and donor templates), and offers potential improvements in efficiency while limiting damage and simplifying the delivery of editing machinery. At the same time, these systems enable novel mutagenesis strategies to introduce sequence diversity for engineering and discovery. Here, we review the different base editing platforms, including their deaminase recruitment strategies and editing outcomes, and compare them to other CRISPR genome editing technologies. Additionally, we discuss how these systems have been applied in therapeutic, engineering, and research settings. Lastly, we explore future directions of this emerging technology.
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