Capturing the dynamic epigenome using single molecule and single cell approaches

摘要： 我们的身体由数十亿个基因相同的细胞组成，这些细胞可以表现出不同的表型或表观遗传学 各州。组蛋白的共价和可逆修饰使细胞能够建立可遗传的基因 在不改变其基因蓝图的情况下，表达模式。控制基因的表观遗传机制 表达对于维持细胞特性和规划多细胞分化是必不可少的。组蛋白H3赖氨酸 9甲基化(H3K9me)与转录沉默和异染色质形成有关。裂解酵母 (S.pombe)有一个最低限度的异染色质结构，它服从于高通量的遗传学和 生物化学。三种保守的蛋白质调节异染色质，包括：1)H3K9me专一性 ‘’编写器，‘’催化H3K9Me的Clr4Suv39h 2)H3K9Me特异的‘’读取器，‘’与H3K9Me结合的Swi6HP1 染色质和沉默转录，以及3)H3K9me特有的“橡皮”，Epe1JmjC，它反对 异染色质组装与表观遗传。CLR4与四环素诱导的TetR DNA融合 结合结构域有助于异染色质组装的快速和可逆控制。我的实验室很有创意 遗传策略使我们能够识别具有独特作用的染色质相关因子，这些因子仅限于 异染色质维持。在补充奖支持的暑期研究体验期间， 这名本科生将专注于利用基于聚合酶链式反应的突变技术产生新的蛋白质变异。 识别增强或抑制表观遗传的等位基因的方法。研究经验是 旨在提升应聘者的技术、操作和专业技能。研究心得 也符合候选人未来的职业抱负，成为生物医学劳动力的一部分，以及 继续研究生教育。此外，这里描述的所有科学目标都与 提出了家长奖的研究方向。

Abstract: Our bodies consist of billions of genetically identical cells that can exhibit distinct phenotypic or epigenetic states. The covalent and reversible modification of histones enables cells to establish heritable gene expression patterns without altering their genetic blueprint. Epigenetic mechanisms that control gene expression are essential to maintain cellular identity and program multicellular differentiation. Histone H3 lysine 9 methylation (H3K9me) is associated with transcription silencing and heterochromatin formation. Fission yeast (S. pombe) has a minimalist heterochromatin architecture that is amenable to high-throughput genetics and biochemistry. A trio of conserved proteins regulates heterochromatin, which includes, 1) an H3K9me specific ''writer,'' Clr4Suv39h that catalyzes H3K9me 2) an H3K9me specific ''reader,'' Swi6HP1 that binds to H3K9me chromatin and silences transcription and, 3) an H3K9me specific ''eraser,'' Epe1JmjC, that opposes heterochromatin assembly and epigenetic inheritance. Fusing Clr4 to the tetracycline-inducible TetR DNA binding domain facilitates rapid and reversible control of heterochromatin assembly. My lab’s innovative genetic strategy has enabled us to identify chromatin-associated factors with unique roles that are restricted to heterochromatin maintenance. During the summer research experience supported by the supplement award, the undergraduate student will focus on generating new protein variants using a PCR based mutagenesis approach to identify alleles that enhance or suppress epigenetic inheritance. The research experience is designed to promote the technical, operation and professional skills of the candidate. The research experience is also aligned with the candidate’s future career aspirations to be a part of the biomedical workforce and pursue a graduate education. Furthermore,all of the scientific goals described here are fully consistent with the proposed research directions of the parent award.