在体内基因组编辑使用金黄色葡萄球菌Cas9
F Ann Ran1, Le Cong2, Winston X Yan3
11] Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA [2] Society of Fellows, Harvard University, Cambridge, Massachusetts 02138, USA.
Nature
|April 2, 2015
概括
较小的金黄色葡萄球菌Cas9 (SaCas9) 能够通过腺相关病毒载体进行高效的基因组编辑. 这种SaCas9系统成功修改了小鼠的Pcsk9基因,降低了高特异性的胆固醇水平.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 遗传学 是一个遗传学.
背景情况:
- Cas9酶是基因组编辑的强大工具.
- 大型的Streptococcus pyogenes Cas9 (SpCas9) 限制了其与腺相关病毒 (AAV) 传递系统的使用.
- 为了有效的AAV介导基因编辑,需要较小的Cas9变体.
研究的目的:
- 为了对更小的Cas9基因组编辑器进行鉴定.
- 评估Staphylococcus aureus Cas9 (SaCas9) 在体内基因编辑的效率和特异性.
- 使用AAV传递来证明SaCas9的治疗潜力.
主要方法:
- 六个较小的Cas9正义词的表征.
- 在小鼠肝脏中进行体内基因组编辑,使用通过AAV传递的SaCas9准Pcsk9基因.
- 评估基因修改效率和降低血清Pcsk9和胆固醇水平.
- 使用BLESS (基于结合和结合的外核酶测序) 进行全基因组特异性分析.
主要成果:
- 黄金葡萄球菌Cas9 (SaCas9) 显示的基因组编辑效率与SpCas9.9相当.
- SaCas9比SpCas9短1千基以上,从而促进了AAV的包装.
- 在体内SaCas9输送导致>40%的Pcsk9基因修饰在小鼠肝脏在一周内.
- 在编辑后观察到血清Pcsk9和总胆固醇水平的显著降低.
- 在体内,SaCas9表现出高效和特定的全基因组向.
结论:
- SaCas9是一个可行的,较小的替代SPCas9用于基因组编辑应用程序.
- 包装在AAV中的SaCas9可实现高效的体内基因编辑,具有治疗潜力.
- 以SaCas9为媒介的编辑为管理诸如高胆固醇血症等疾病提供了一个有希望的策略.
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