近期《自然—方法學(xué)》報(bào)道了如何利用特定核酸酶對(duì)導(dǎo)致瘧疾發(fā)病的惡性瘧原蟲的基因組進(jìn)行修改。這項(xiàng)發(fā)現(xiàn)將有助于從遺傳層面包括抗藥性機(jī)制上更深入地研究這種重要的人體寄生蟲。
全球有數(shù)以百萬計(jì)的人飽受瘧疾侵害,，而抗藥性是瘧疾治療過程中普遍存在的問題,。針對(duì)引發(fā)瘧疾的原生動(dòng)物寄生蟲的研究受到阻礙,，原因在于缺乏耐用的研究工具，特別是那些能操控這些寄生蟲基因組的工具,。
鋅指核酸酶（ZFN）可被設(shè)計(jì)用來切入各種基因組的目標(biāo)位置并使其發(fā)生改變,。已有科學(xué)家將其用于修改蠅類、魚類,、蠕蟲,、人體細(xì)胞等各種生物類型的基因組。
David Fidock等人在文章中報(bào)告了如何利用這類工具修改惡性瘧原蟲這種最致命瘧疾寄生蟲的基因組,。該寄生蟲基因組中核苷酸的平衡量及其內(nèi)生DNA修復(fù)的機(jī)械性因素中的某些方面與其他有機(jī)體有所差別,，其中，DNA修復(fù)的機(jī)械性因素本可能削弱ZFN的作用效果,。但研究人員展示了他們利用該工具對(duì)惡性瘧原蟲基因組有效的操控,，他們使用ZFN刪除一些基因、替換掉一個(gè)基因,，以快速而有效的方式改變了其基因序列,。（生物谷Bioon.com）
doi:10.1038/nmeth.2143
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Site-specific genome editing in Plasmodium falciparum using engineered zinc-finger nucleases
Judith Straimer,Marcus C S Lee,Andrew H Lee,Bryan Zeitler,April E Williams,Jocelynn R Pearl,Lei Zhang,Edward J Rebar,Philip D Gregory,Manuel Llinás,Fyodor D Urnov & David A Fidock
Malaria afflicts over 200 million people worldwide, and its most lethal etiologic agent, Plasmodium falciparum, is evolving to resist even the latest-generation therapeutics. Efficient tools for genome-directed investigations of P. falciparum-induced pathogenesis, including drug-resistance mechanisms, are clearly required. Here we report rapid and targeted genetic engineering of this parasite using zinc-finger nucleases (ZFNs) that produce a double-strand break in a user-defined locus and trigger homology-directed repair. Targeting an integrated egfp locus, we obtained gene-deletion parasites with unprecedented speed (2 weeks), both with and without direct selection. ZFNs engineered against the parasite gene pfcrt, responsible for escape under chloroquine treatment, rapidly produced parasites that carried either an allelic replacement or a panel of specified point mutations. This method will enable a diverse array of genome-editing approaches to interrogate this human pathogen.
