TitleOptimizing the detection of nascent transcripts by RNA fluorescence in situ hybridization.
Publication TypeJournal Article
Year of Publication2001
Authorsvan Raamsdonk, CD, Tilghman, SM
JournalNucleic Acids Res
Date Published2001 Apr 15
KeywordsAlleles, Animals, Cell Nucleus, Cells, Cultured, DNA, Eye Proteins, Fluorescent Dyes, Gene Expression Regulation, Developmental, Homeodomain Proteins, In Situ Hybridization, Fluorescence, Mice, Paired Box Transcription Factors, Repressor Proteins, Retinal Ganglion Cells, RNA, Messenger, Sensitivity and Specificity, Transcription, Genetic

An unusual feature of the mammalian genome is the number of genes exhibiting monoallelic expression. Recently random monoallelic expression of autosomal genes has been reported for olfactory and Ly-49 NK receptor genes, as well as for Il-2, Il-4 and Pax5. RNA fluorescence in situ hybridization (FISH) has been exploited to monitor allelic expression by visualizing the number of sites of transcription in individual nuclei. However, the sensitivity of this technique is difficult to determine for a given gene. We show that by combining DNA and RNA FISH it is possible to control for the hybridization efficiency and the accessibility and visibility of fluorescent probes within the nucleus.

Alternate JournalNucleic Acids Res.