Abstract

During fiscal year 2011, we accomplished the following: 1. We identified homologous recombinants amongst greater than 200 embryonic stem (ES) cell clones using Southern blotting and PCR assays. These ES cells were generated using targeting vectors that introduced LacO or TetO binding-sites in the 1) VH locus, 2) 3 regulatory region, 3) DH locus, and 4) within the intronic enhancer, E. The identified clones are being used for embryo reconstitution to generate targeted mice. 2. We developed stable cell lines in mouse embryo fibroblasts (MEFs) using two manipulated bacterial artificial chromosomes (BACs). Both BACs are derived from the VH region of the murine IgH locus but differ in the number of naturally occurring L1 retrotransposons. We will use these cell lines to determine if L1 retrotransposons can initiate heterochromatinization and drive associated sequences to the nuclear periphery.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAAG000372-04
Application #
8335842
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2011
Total Cost
$445,646
Indirect Cost

  Institution

Name
National Institute on Aging
Department
Type
DUNS #
City
State
Country
Zip Code

  Publications

Qiu, Xiang; Kumari, Gita; Gerasimova, Tatiana et al. (2018) Sequential Enhancer Sequestration Dysregulates Recombination Center Formation at the IgH Locus. Mol Cell 70:21-33.e6
Montefiori, Lindsey; Wuerffel, Robert; Roqueiro, Damian et al. (2016) Extremely Long-Range Chromatin Loops Link Topological Domains to Facilitate a Diverse Antibody Repertoire. Cell Rep 14:896-906
Gerasimova, Tatiana; Guo, Changying; Ghosh, Amalendu et al. (2015) A structural hierarchy mediated by multiple nuclear factors establishes IgH locus conformation. Genes Dev 29:1683-95
Phillips-Cremins, Jennifer E; Sauria, Michael E G; Sanyal, Amartya et al. (2013) Architectural protein subclasses shape 3D organization of genomes during lineage commitment. Cell 153:1281-95
Selimyan, Roza; Gerstein, Rachel M; Ivanova, Irina et al. (2013) Localized DNA demethylation at recombination intermediates during immunoglobulin heavy chain gene assembly. PLoS Biol 11:e1001475
Kumar, Satyendra; Wuerffel, Robert; Achour, Ikbel et al. (2013) Flexible ordering of antibody class switch and V(D)J joining during B-cell ontogeny. Genes Dev 27:2439-44
Subrahmanyam, Ramesh; Sen, Ranjan (2012) Epigenetic features that regulate IgH locus recombination and expression. Curr Top Microbiol Immunol 356:39-63
Subrahmanyam, Ramesh; Du, Hansen; Ivanova, Irina et al. (2012) Localized epigenetic changes induced by DH recombination restricts recombinase to DJH junctions. Nat Immunol 13:1205-12
Roy, Ananda L; Sen, Ranjan; Roeder, Robert G (2011) Enhancer-promoter communication and transcriptional regulation of Igh. Trends Immunol 32:532-9
Osipovich, Oleg A; Subrahmanyam, Ramesh; Pierce, Steven et al. (2009) Cutting edge: SWI/SNF mediates antisense Igh transcription and locus-wide accessibility in B cell precursors. J Immunol 183:1509-13

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