Epigenetic editing by CRISPR/dCas9 in Plasmodium falciparum
Epigenetic editing by CRISPR/dCas9 in Plasmodium falciparum
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恶性疟原虫中的 CRISPR/dCas9 表观遗传编辑
DOI：
10.1073/pnas.1813542116
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Jiang Lubin
中科院分区：
文献类型：
--
作者：
Xiao Bo;Yin Shigang;Hu Yang;Sun Maoxin;Wei Jieqiong;Huang Zhenghui;Wen Yuhao;Dai Xueyu;Chen Huiling;Mu Jianbing;Cui Liwang;Jiang Lubin
由于恶性疟原虫基因组中同源重组效率低且缺乏RNAi，因此迫切需要可行的基因编辑工具包。尽管各种基因编辑策略已成功应用于恶性疟原虫，但仍存在严重的局限性。在这里，我们表明，通过将组蛋白乙酰转移酶或脱乙酰酶融合到CRISPR/dCas 9（成簇规则间隔短回文重复序列/CRISPR相关蛋白9）系统中的dCas 9突变体，可以在恶性疟原虫中有效特异地调控靶基因的表达。这两个表观遗传学工具包将扩大目前恶性疟原虫功能遗传学和表观遗传学研究的应用，从而加速对疟疾发病机制的理解。遗传操作仍然是了解最致命的疟疾寄生虫恶性疟原虫的功能基因组学的主要障碍。尽管CRISPR/Cas9（成簇规则间隔短回文重复序列/CRISPR相关蛋白9）系统已成功应用于在寄生虫基因组中引入永久性变化，但其使用仍然有限。在这里，我们表明，将不同的表观遗传效应子结构域融合到Cas9无效突变体有效地并且特异性地重新编程恶性疟原虫中靶基因的表达。通过分别精确地写入和擦除侵入相关基因网织红细胞结合蛋白同源物4（rh 4）和红细胞结合蛋白175（eba-175）转录起始位点区域的组蛋白乙酰化，我们实现了rh 4的显著激活和eba-175的抑制，导致寄生虫侵入途径切换到人类红细胞。通过使用表观遗传敲低系统，我们还表征了PfSET 1的作用，PfSET 1先前被鉴定为必需基因，对主要滋养体和寄生虫特异性基因的表达，因此调节恶性疟原虫成熟形式的生长。这种表观遗传CRISPR/dCas 9系统为在恶性疟原虫中在转录水平上调节基因表达提供了有力的方法。
Significance As a result of low homologous recombination efficiency and the lack of RNAi in the Plasmodium falciparum genome, feasible gene-editing toolkits are urgently needed. Although various gene editing strategies have been successfully applied to P. falciparum, there are still serious limitations. Here we show that by fusing histone acetyltransferase or deacetylase to a dCas9 mutant in the CRISPR/dCas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) system, the expression of target genes could be efficiently and specifically regulated in P. falciparum. The two epigenetic toolkits will expand current applications for functional genetics and epigenetics studies in P. falciparum, and therefore accelerate the understanding of malaria pathogenesis. Genetic manipulation remains a major obstacle for understanding the functional genomics of the deadliest malaria parasite Plasmodium falciparum. Although the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) system has been successfully applied to introduce permanent changes in the parasite genome, its use is still limited. Here we show that fusing different epigenetic effector domains to a Cas9 null mutant efficiently and specifically reprograms the expression of target genes in P. falciparum. By precisely writing and erasing histone acetylation at the transcription start site regions of the invasion-related genes reticulocyte binding protein homolog 4 (rh4) and erythrocyte binding protein 175 (eba-175), respectively, we achieved significant activation of rh4 and repression of eba-175, leading to the switch of the parasite invasion pathways into human erythrocytes. By using the epigenetic knockdown system, we have also characterized the effects of PfSET1, previously identified as an essential gene, on expression of mainly trophozoite- and schizont-specific genes, and therefore regulation of the growth of the mature forms of P. falciparum. This epigenetic CRISPR/dCas9 system provides a powerful approach for regulating gene expression at the transcriptional level in P. falciparum.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
48
作者：
Wagner, Jeffrey C.;Platt, Randall J.;Goldfless, Stephen J.;Zhang, Feng;Niles, Jacquin C.
通讯作者：
Niles, Jacquin C.
DOI：
--
发表时间：
1995
期刊：
--
影响因子：
--
作者：
Y. Wu
通讯作者：
Y. Wu
影响因子：
--
作者：
Merrick, Catherine J.;Duraisingh, Manoj T.
通讯作者：
Duraisingh, Manoj T.
DOI：
10.1073/pnas.0504679102
发表时间：
2005-11-08
影响因子：
11.1
作者：
Balu, B;Shoue, DA;Adams, JH
通讯作者：
Adams, JH