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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jul 16, 2008 |
| Title |
Gene expression changes in the intestinal crypts of mice with loss of imprinting |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
To identify genes differentially expressed in the intestinal crypts between LOI(+) and LOI(-) mice, we performed microarray experiments using RNA extracted from laser capture microdissection, by microdissecting an average of 8,000 crypts from each of 3 LOI(+) and 3 LOI(-) mice.
Keywords: genetic modification design
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| Overall design |
H19 mutant mice with C57BL/6J background carrying a deletion in the H19 gene (3 kb) and 10 kb of the upstream region including differentially methylated region (DMR) were maintained without LOI phenotype by breeding female wild-type C57BL/6J and male H19+/-. Experimental crosses were performed between female H19+/- and male wild-type C57BL/6J to obtain LOI(+) mice and LOI(-) mice. Igf2 on the maternal allele is silenced in LOI(-), i.e. wild type, mice. But when H19 DMR deletion is inherited from mother, the normally silenced Igf2 on maternal allele is activated and LOI (loss of imprinting) of Igf2 occurs. LOI(+) and LOI(-) mice were sacrificed at 120 days, and the tissue pieces of the small intestine were collected from the middle (the third fifth) part and kept frozen at -80C. To detect gene expression change in intestinal progenitor cells clearly, Laser Capture Microdissection (LCM) was performed to collect intestinal crypt cells. The 10 um thick sections were prepared from the frozen small intestine pieces, and 8,000 intestinal crypts at average were dissected by LCM for each of three LOI(+) and LOI(-) mice, and 2 ug of total RNA at least were collected using RNeasy Kit (GIAGEN). 1.7 ug of total RNA for each sample were used for labeling to compare gene expressions in intestinal crypt between LOI(+) and LOI(-) mice.
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| Contributor(s) |
Kaneda A, Wang CJ, Cheong R, Onyango P, Wen B, Iacobuzio-Donahue CA, Ohlsson R, Andraos R, Pearson MA, Sharov AA, Longo DL, Ko MS, Levchenko A, Feinberg AP |
| Citation(s) |
18087038 |
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| Submission date |
Jul 25, 2007 |
| Last update date |
Jan 17, 2013 |
| Contact name |
Andrew Feinberg |
| E-mail(s) |
afeinberg@jhu.edu
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| Phone |
4106143489
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| Fax |
4106149819
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| Organization name |
Johns Hopkins University
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| Department |
Medicine
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| Lab |
Center for Epigenetics
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| Street address |
720 Rutland Ave., 1064 Ross
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| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
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| Platforms (1) |
| GPL2552 |
NIA Mouse 44K Microarray v2.1 (Whole Genome 60-mer Oligo) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA101733 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE8583_RAW.tar |
200.8 Mb |
(http)(custom) |
TAR (of TIFF, TXT) |
| Processed data included within Sample table |
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