Activation and inactivation of VH gene recombination

VH基因重组的激活和失活

基本信息

批准号：
7963946
负责人：
RANJAN SEN
金额：
$ 33.08万
依托单位：
NATIONAL INSTITUTE ON AGING
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7963946
关键词：
Antigen Receptors B-Lymphocytes Bone Marrow Boundary Elements Chromosomes, Human, Pair 12 Development Employee Strikes Epigenetic Process Exclusion Functional RNA Gene Rearrangement Gene Silencing Genes Genetic Recombination Genetic Transcription Goals IGH@ gene cluster L1 Elements Location Maps Molecular Mus Positioning Attribute Process Receptor Gene Rest Role Staging Standardization Sterility Technology Tetanus Helper Peptide chromatin modification chromosome movement design telomere vector

Antigen Receptors B-Lymphocytes Bone Marrow Boundary Elements Chromosomes, Human, Pair 12 Development Employee Strikes Epigenetic Process Exclusion Functional RNA Gene Rearrangement Gene Silencing Genes Genetic Recombination Genetic Transcription Goals IGH@ gene cluster L1 Elements Location Maps Molecular Mus Positioning Attribute Process Receptor Gene Rest Role Staging Standardization Sterility Technology Tetanus Helper Peptide chromatin modification chromosome movement design telomere vector

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项目摘要

During the 2009 fiscal year we accomplished the following: 1. Continued studies of sterile transcription and epigenetic state of unrearranged VH gene segments in pro-B cells from RAG2-deficient bone marrow. We mapped the position and distribution of L1 retrotransposous in the VH locus, and found one striking cluster of 4 L1 elements that separates the 5-prime most 5% VH gene segments from the rest of the VH genes. We propose that this cluster of L1s may serve as a boundary element to delineate independently regulated VH domains. 2. Designed additional primer pairs to score for transcription and chromatin modifications in the sub-telomeric region of mouse chromosome 12. Standardization of these newly-designed primers is ongoing. 3. Constructed a targeting vector to replace endogenous E with multimerized Gal4 and Tet-operator sequences. Used BAC recombineering technology to construct targeting vectors to introduce Gal4, Tet0, and Lac0 sequences at different locations of the VH locus. These vectors will be used to alter the IgH locus in order to introduce a tag that can be used to visualize chromosome movements during antigen receptor gene rearrangements.

在2009财政年度，我们完成了以下内容： 1。持续研究来自RAG2缺陷型骨髓的pro-B细胞中透明的VH基因段的无菌转录和表观遗传态。我们绘制了L1逆转录座位在VH基因座中的位置和分布，并找到了一个4个L1元素的引人注目的群集，该集群将5％的5％VH基因段与其他VH基因分开。我们建议该L1群可以用作划定独立调节VH域的边界元素。 2。设计了其他引物对以在小鼠染色体12的亚层次区域进行转录和染色质修饰。这些新设计的引物的标准化正在进行中。 3。构建了一个靶向载体，用多层化的GAL4和TET-ESTERATOR序列代替内源E。使用BAC重组技术来构建靶向向量，以在VH基因座的不同位置引入GAL4，TET0和LAC0序列。这些向量将用于改变IGH基因座，以引入一个标签，该标签可用于在抗原受体基因重排期间可视化染色体运动。