Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011:676:85-100.
doi: 10.1007/978-1-60761-863-8_7.

MicroRNA Northern blotting, precursor cloning, and Ago2-improved RNA interference

Julia Winter  1 , Sven Diederichs
Affiliations

MicroRNA Northern blotting, precursor cloning, and Ago2-improved RNA interference

Julia Winter et al. Methods Mol Biol. 2011.

Abstract

Aberrant expression of microRNAs (miRNAs) and processing defects in their biogenesis pathway are a widespread phenomenon in tumors, conveying great importance to the analysis of miRNA expression, regulation, and biogenesis to gain knowledge about their role in cancer. Besides Drosha and Dicer, Argonaute proteins are key players in miRNA processing. In addition to their role as components of the RNA-induced silencing complex (RISC) executing target silencing, Argonautes mediate posttranscriptional regulation of miRNA maturation by creating an additional intermediate processing step, the Ago2-cleaved precursor miRNA (ac-pre-miRNA), and enhancing the production or stability of mature miRNAs. Here, we describe the detection of miRNA levels by Northern blotting and the identification of the 3' end of miRNAs by precursor cloning to accentuate two of the many roles of Argonaute proteins. In addition, we describe a method to optimize RNAi experiments by increasing the efficacy and specificity of target silencing via Ago2 cotransfection.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

LinkOut - more resources