The goal of this project is to develop a method to monitor the expression levels of a large number of genes in various experimental conditions and to elucidate the global structure and behavior of a gene regulatory network in development and aging. In our previous work, we have constructed cDNA libraries from early mouse embryos and stem cells and generated a large number of expressed sequence tags (ESTs). During the last one-year period, we have accomplished the following additional goals. 1) We have developed a glass-slide microarray platform containing in situ-synthesized 60-mer oligonucleotide probes representing approximately 44,000 unique mouse transcripts. 2) We have produced web-based ANOVA-FDR software to provide user-friendly microarray data analysis too. 3) We have developed an algorithm and a fully-automated computational pipeline for transcript assembly from expressed sequences aligned to the mouse genome. We have identified 191,946 genomic loci, which included 27,497 protein-coding genes and 11,906 additional gene candidates (e.g., non-protein-coding, but multi-exon). 4) We have developed a high throughput whole-mount in situ hybridization technique for preimplantation mouse embryos and ES cells. The nonrestricted community access to the resource can accelerate a wide range of research, particularly in reproductive and regenerative medicine.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Intramural Research (Z01)
Project #
1Z01AG000656-07
Application #
7132304
Study Section
(LG)
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2005
Total Cost
Indirect Cost
Name
Aging
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Sharov, Alexei A (2017) Composite Agency: Semiotics of Modularity and Guiding Interactions. Biosemiotics 10:157-178
Sharov, Alexei A (2016) Coenzyme world model of the origin of life. Biosystems 144:8-17
Sharov, Alexei A (2016) Evolutionary biosemiotics and multilevel construction networks. Biosemiotics 9:399-416
Teratani-Ota, Yusuke; Yamamizu, Kohei; Piao, Yulan et al. (2016) Induction of specific neuron types by overexpression of single transcription factors. In Vitro Cell Dev Biol Anim 52:961-973
Livigni, Alessandra; Peradziryi, Hanna; Sharov, Alexei A et al. (2013) A conserved Oct4/POUV-dependent network links adhesion and migration to progenitor maintenance. Curr Biol 23:2233-2244
Morgani, Sophie M; Canham, Maurice A; Nichols, Jennifer et al. (2013) Totipotent embryonic stem cells arise in ground-state culture conditions. Cell Rep 3:1945-57
Hammachi, Fella; Morrison, Gillian M; Sharov, Alexei A et al. (2012) Transcriptional activation by Oct4 is sufficient for the maintenance and induction of pluripotency. Cell Rep 1:99-109
Canham, Maurice A; Sharov, Alexei A; Ko, Minoru S H et al. (2010) Functional heterogeneity of embryonic stem cells revealed through translational amplification of an early endodermal transcript. PLoS Biol 8:e1000379
Kunisada, Makoto; Cui, Chang-Yi; Piao, Yulan et al. (2009) Requirement for Shh and Fox family genes at different stages in sweat gland development. Hum Mol Genet 18:1769-78
Sharova, Lioudmila V; Sharov, Alexei A; Nedorezov, Timur et al. (2009) Database for mRNA half-life of 19 977 genes obtained by DNA microarray analysis of pluripotent and differentiating mouse embryonic stem cells. DNA Res 16:45-58

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