FUNCTIONAL ANALYSIS OF IMPRINTING MUTATIONS

描述：（改编自申请人的摘要）： 染色体15 q11-q13中的印记过程导致 在亲本配子发生期间，用Angelman或 Prader-Willi综合征（AS或PWS），取决于父母的起源 变异的原因 AS印记突变患者的基因突变， 元素的PI。提示对照组父系至母系（pat >mat）印记 在女性生殖细胞中转换，而PWS家族在 SNRPN启动子，他建议控制mat >pat印记转换， 雄性生殖系 这些AS和PWS突变定义了一个假定的印记 中心（IC）。 这里提出的研究有三个具体的目标来测试 这些假设，通过制作印迹突变的小鼠模型：（1） IC转录物的分离和表征（pat >mat开关元件， XX种系）和Snrpn启动子（mat >pat开关元件，XY种系）， 测试这些元素在印记转换中的作用;（2）目标 通过胚胎干中的同源重组将突变引入小鼠IC 细胞，以最终测试这些元素的假设作用， 印记转换;（3）检查印记基因的表观基因型结果 在IC控制的2 Mb结构域中，在表达和DNA甲基化时， 从遗传的印记突变，并检查表型 方面的影响.

DESCRIPTION: (adapted from Applicant's Abstract): Mutations in the imprinting process in chromosome 15q11-q13 cause failure to switch the imprint of a 2 Mb region during parental gametogenesis, with Angelman or Prader-Willi syndrome (AS or PWS) arising, depending on the parental origin of the mutation. AS imprinting mutation patients have mutations in an element the P.I. suggests controls paternal to maternal (pat >mat) imprint switching in the female germline, while PWS families have mutations in the SNRPN promoter, which he suggests controls mat >pat imprint switching in the male germline. These AS and PWS mutations define a putative imprinting center (IC). The studies proposed here have three specific aims to test these hypotheses, by making mouse models of imprinting mutations: (1) Isolation and characterization of IC transcripts (pat >mat switch element, XX germline) and the Snrpn promoter (mat >pat switch element, XY germline), to test the role of these elements in imprint switching; (2) Target mutations into the mouse IC by homologous recombination in embryonic stem cells, to conclusively test the role hypothesized for these elements in imprint switching; (3) Examine the epigenotypic outcome for imprinted genes in the 2 Mb domain controlled by the IC, on expression and DNA methylation, from inheritance of imprinting mutations, and examine for phenotypic effects.