新型DNA表观遗传修饰检测技术的开发及在体细胞重编程研究中的应用

Environmental and genetic factors dynamically regulate gene expression through DNA modifications exemplified by 5-methylcytosine, thereby determining cell fate. A key bottleneck to the study of these DNA modifications is the development of robust sequencing technologies towards such epigenetic modifications. Yet, unsovled limitations of the existing technologies have significantly impeded our understanding to the detecion and functional investigations of these novel modifications. Here, we propose the development of several novel sequencing technologies for the systematic detection of 5-hydroxylmethylcytosine, 5-formylcytosine, 5-carboxylcytosine, and 6-methyladenosine. Such methods will also be applied to reveal the comprehensive spectrum of these DNA modifications (5-methylcytosine, 5-hydroxylmethylcytosine, 5-formylcytosine and 5-carboxylcytosine) during somatic cell reprogramming; and the relationship between environmental factors (such as small molecules) and the DNA modifications will be profiled as well. Finally, both the sequencing and detection technologies and the dynamic modification spetrum generated by this study can be shared and applied by researchers to study other questions in cell prgramming and reprogramming.

环境与遗传因素通过以DNA甲基化为代表的表观遗传修饰，动态调控基因表达，从而影响细胞命运。DNA修饰测序技术的发展，为新颖DNA修饰的功能研究奠定了基础。然而，现有测序技术存在明显不足，严重影响了新颖DNA修饰的检测与功能研究。本项目将开发DNA修饰（5-羟甲基胞嘧啶、5-醛基胞嘧啶、5-羧基胞嘧啶及6-甲基腺嘌呤）的新型检测技术，系统研究各种DNA修饰在体细胞重编程过程中的全谱系变化（5-甲基胞嘧啶、5-强甲基胞嘧啶、5-醛基胞嘧啶和5-羧基胞嘧啶），探究小分子及环境因素对DNA修饰谱系的影响。本项目开发的DNA修饰测序与检测技术、产生的DNA修饰谱系动态关联数据，也有望应用于细胞编程与重编程领域内其他以DNA甲基化为代表的表观遗传修饰研究。