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普渡大学团队发现 NgAgo 基因编辑功能

已有 4128 次阅读 2019-5-11 13:16 |个人分类:生物|系统分类:海外观察

怎么看?我之前在《论对韩春雨的彻底否定》一文中写道:【NgAgo 既然已经被独立确定具备定点切割 RNA 的功能,能否在其基础上进行改造优化,使之具备对 DNA 断链的能力? 我不认为这个问题谁能用薛定谔方程证明为“不可能”。所以,不应该对韩春雨彻底 否定,而是要看到差距,而是应该继续研究。古书上断然写着青蒿能治疟疾,实际喝青蒿水有效率几乎为零,过了一千多年屠呦呦却拿了个诺贝尔奖。

翻译:普渡大学的研究小组创建了一种方法用以DNA为导引 的 NgAgo 在任意点进行基因编辑。


The Purdue team created a method that uses the protein Argonaute from Natronobacterium gregoryi (NgAgo) and supplied DNA as a guide to enable modification anywhere on the genome, providing new options to potentially improve manufacturing, disease treatment, drug discovery and crop production.

论文网址: https://www.biorxiv.org/content/10.1101/597237v1

NgAgo is a DNA-guided DNA endonuclease Although previous studies demonstrated that refolded NgAgo does not cut DNA in vitro , consistent with our findings, we establish that soluble NgAgo can, in fact, cleave DNA in vitro. That is, refolded NgAgo may not be fully functional. As we showed that an N-del/D663A/D738A catalytic mutant lacks DNA cleaving activity (Fig. 2e), the catalytic activity is unlikely to be the result of sample contamination. However, we are unable to demonstrate unequivocal guide-dependent cleavage with both double-stranded DNA target and  single-stranded DNA target in vitro (data not shown). This may be due to inefficient guide loading, as we observe that N-del co-purifies guides (Fig. 2c). 

NgAgo can be repurposed as a DNA editing tool 

Our results provide supporting evidence to encourage the development of NgAgo for gene-editing.


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