期刊
METHODS
卷 43, 期 2, 页码 153-161出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2007.04.008
关键词
MicroRNAs; gene expression; nervous system; brain; development; embryos; fluorescein; mouse; rat; human
资金
- NICHD NIH HHS [R03 HD043828-02, R03 HD043828, R03 HD043828-02S1] Funding Source: Medline
- NIDDK NIH HHS [R21 DK068059-02, R21 DK068059-01, R21 DK068059] Funding Source: Medline
- NIMH NIH HHS [P50 MH60398] Funding Source: Medline
In situ hybridization is an important tool for analyzing gene expression and developing hypotheses about gene functions. The discovery of hundreds of microRNA (miRNA) genes in animals has provided new challenges for analyzing gene expression and functions. The small size of the mature miRNAs (similar to 20-24 nucleotides in length) presents difficulties for conventional in situ hybridization methods. However, we have described a modified in situ hybridization method for detection of mammalian miRNAs in tissue sections, based upon the use of RNA oligonucleotide probes in combination with highly specific wash conditions. Here, we present detailed procedures for detection of miRNAs in tissue sections or cultured cells. The methods described can utilize either nonradioactive hapten-conjugated probes that are detected by enzyme-coupled antibodies, or radioactively labeled probes that are detected by auto radiography. The ability to visualize miRNA expression patterns in tissue sections provides an additional tool for the analyses of miRNA expression and function. In addition, the use of radioactively labeled probes should facilitate quantitative analyses of changes in miRNA gene expression. (C) 2007 Elsevier Inc. All rights reserved.
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