TY - JOUR
T1 - Optimizing the detection of nascent transcripts by RNA fluorescence in situ hybridization
AU - Van Raamsdonk, Catherine D.
AU - Tilghman, Shirley M.
N1 - Funding Information:
We would like to thank Dr Barbara Panning, now at the University of California San Francisco, for her RNA FISH protocol. S.M.T. is an Investigator of the Howard Hughes Medical Institute, which supported this work.
Publisher Copyright:
© 2001 Oxford University Press.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/29.8.e42
DO - 10.1093/nar/29.8.e42
M3 - Article
C2 - 11292856
AN - SCOPUS:0035872080
SN - 0305-1048
VL - 29
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 8
M1 - e42
ER -