Optimization of CRISPR/Cas System for Improving Genome Editing Efficiency in Plasmodium falciparum.
Optimization of CRISPR/Cas System for Improving Genome Editing Efficiency in Plasmodium falciparum.
复制标题
优化 CRISPR/Cas 系统以提高恶性疟原虫基因组编辑效率。
DOI：
10.3389/fmicb.2020.625862
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发表时间：
2020
影响因子：
5.2
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Zhao Y;Wang F;Wang C;Zhang X;Jiang C;Ding F;Shen L;Zhang Q
长期以来，疟疾寄生虫的分子机制和相关基因功能的研究受到有限的基因组编辑技术的限制。最近，一种简单而有效的基因组编辑技术，CRISPR/Cas（clustered regularly interspaced short palindromic repeats/CRISPR associated）系统，极大地促进了许多生物体（包括疟疾寄生虫）的这些研究。然而，由于疟疾寄生虫的特殊基因组特征，CRISPR/Cas系统在这种病原体中的操作和基因编辑功效需要改进，特别是在人类疟疾寄生虫恶性疟原虫中。在此，基于CRISPR/Cas9系统，我们开发了一种整合策略来产生Cas9 i系统，其显著缩短了在恶性疟原虫中产生转基因菌株的时间。此外，利用这种Cas9 i系统，我们已经成功地通过恶性疟原虫中的单轮转染实现了多重基因组编辑（突变或标记）。此外，我们首次将Cas9的替代品AsCpf 1（Acidaminococcus sp.Cpf1）适应于恶性疟原虫寄生虫，并对其进行基因编辑。CRISPR/Cas系统的这些优化将进一步促进疟疾寄生虫的机制研究，并有助于未来消除疟疾。
Studies of molecular mechanisms and related gene functions have long been restricted by limited genome editing technologies in malaria parasites. Recently, a simple and effective genome editing technology, the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system, has greatly facilitated these studies in many organisms, including malaria parasites. However, due to the special genome feature of malaria parasites, the manipulation and gene editing efficacy of the CRISPR/Cas system in this pathogen need to be improved, particularly in the human malaria parasite, Plasmodium falciparum. Herein, based on the CRISPR/Cas9 system, we developed an integrating strategy to generate a Cas9i system, which significantly shortened the time for generation of transgenic strains in P. falciparum. Moreover, with this Cas9i system, we have successfully achieved multiplexed genome editing (mutating or tagging) by a single-round transfection in P. falciparum. In addition, we for the first time adapted AsCpf1 (Acidaminococcus sp. Cpf1), an alternative to Cas9, into P. falciparum parasites and examined it for gene editing. These optimizations of the CRISPR/Cas system will further facilitate the mechanistic research of malaria parasites and contribute to eliminating malaria in the future.
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影响因子：
48
作者：
Mali, Prashant;Esvelt, Kevin M.;Church, George M.
通讯作者：
Church, George M.
影响因子：
1.5
作者：
Ganesan, Suresh M.;Morrisey, Joanne M.;Ke, Hangjun;Painter, Heather J.;Laroiya, Kamal;Phillips, Margaret A.;Rathod, Pradipsinh K.;Mather, Michael W.;Vaidya, Akhil B.
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Vaidya, Akhil B.
影响因子：
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作者：
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通讯作者：
Niles, Jacquin C.
影响因子：
64.8
作者：
通讯作者：
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影响因子：
64.8
作者：
Komor AC;Kim YB;Packer MS;Zuris JA;Liu DR
通讯作者：
Liu DR