编辑哺乳动物基因组中的DNA甲基化
X Shawn Liu1, Hao Wu1, Xiong Ji1
1Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
Cell
|September 24, 2016
概括
研究人员使用dCas9与Tet1或Dnmt3a融合开发了新的DNA甲基化编辑工具. 这些表观遗传编辑工具通过向DNA甲基化来精确控制基因表达,在小鼠中已经得到了应用.
科学领域:
- 表观遗传学和分子生物学
- 基因调控
- 基因组工程
背景情况:
- 哺乳动物DNA甲基化是调节基因表达的关键表观遗传机制.
- 研究特定DNA甲基化位点的功能影响存在重大挑战.
研究的目的:
- 开发和验证针对性的DNA甲基化编辑系统.
- 证明精确改变DNA甲基化模式及其对基因表达的下游影响的能力.
主要方法:
- 催化无效的Cas9 (dCas9) 与Tet1或Dnmt3a酶的融合
- 针对dCas9-Tet1或dCas9-Dnmt3a特定的基因组位置,包括促进剂和增强剂.
- 评估针对性编辑的DNA甲基化,基因表达和蛋白质结合的变化.
主要成果:
- 对dCas9-Tet1或dCas9-Dnmt3a的向输送使得DNA分别可以进行精确的脱甲基化或甲基化.
- 在神经元中通过dCas9- Tet1调解BDNF表达和MyoD激活以进行细胞重编程.
- 通过干扰CTCF结合和DNA循环,dCas9-Dnmt3a改变了基因表达.
结论:
- dCas9融合系统提供了针对性DNA甲基化编辑的强大平台.
- 这些工具有助于在各种生物环境中对表观遗传调节的功能研究,包括体内小鼠模型.
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