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Revealing details: whole mount microRNA in situ hybridization protocol for zebrafish embryos and adult tissues.

ABSTRACT: Non-coding microRNA (miRNA) molecules bind their target mRNAs and thereby modulate the amount of protein produced. To understand the significance of a potential miRNA-mRNA interaction, temporal and spatial information on miRNA and mRNA expression is essential. Here, we provide a detailed protocol for miRNA whole mount in situ hybridization. We introduce the use of Morpholino based oligos as antisense probes for miRNA detection, in addition to the current "gold standard" locked nucleic acid (LNA) probes. Furthermore we have modified existing miRNA in situ protocols thereby improving both sensitivity and resolution of miRNA visualization in whole zebrafish embryos and adult tissues.

SUBMITTER: Lagendijk AK

PROVIDER: S-EPMC3509442 | biostudies-other | 2012 Jun

REPOSITORIES: biostudies-other

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Revealing details: whole mount microRNA in situ hybridization protocol for zebrafish embryos and adult tissues.

Lagendijk Anne Karine AK Moulton Jon D JD Bakkers Jeroen J

Biology open 20120411 6

Non-coding microRNA (miRNA) molecules bind their target mRNAs and thereby modulate the amount of protein produced. To understand the significance of a potential miRNA-mRNA interaction, temporal and spatial information on miRNA and mRNA expression is essential. Here, we provide a detailed protocol for miRNA whole mount in situ hybridization. We introduce the use of Morpholino based oligos as antisense probes for miRNA detection, in addition to the current "gold standard" locked nucleic acid (LNA) ...[more]

PMID: 23213449

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