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Updated: May 19, 2026
10:53
Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
Published on: January 16, 2017
Co-Immunoprecipitation of long noncoding RNAs
Victoria A Moran1, Courtney N Niland, Ahmad M Khalil
1Department of Genetics, Center for RNA Molecular Biology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 22, 2012
Summary
Researchers explore RIP-Seq, a method to identify long noncoding RNAs (lncRNAs) interacting with proteins. This technique aids in understanding lncRNA functions in gene regulation and diseases like cancer.
Area of Science:
- Molecular Biology
- Genomics
- Biochemistry
Background:
- The human genome encodes thousands of long noncoding RNAs (lncRNAs).
- lncRNAs play crucial roles in biological functions, including gene expression regulation.
- Dysregulation of lncRNAs is linked to human diseases such as cancer and neurological disorders.
Purpose of the Study:
- To address the growing need for methods to elucidate lncRNA functions and mechanisms.
- To introduce and discuss RIP-Seq as a technique for discovering protein-lncRNA interactions.
Main Methods:
- RIP-Seq involves immunoprecipitation of a target protein from cross-linked cell lysates.
- Subsequent steps include reverse-cross-linking, RNA isolation, and deep sequencing.
- This enables the identification of lncRNAs associated with the specific protein complex.
Main Results:
- The study details the RIP-Seq procedure for identifying lncRNA partners of proteins.
- This method facilitates the discovery of novel functional associations for lncRNAs.
Conclusions:
- RIP-Seq is a valuable method for investigating the functional roles of lncRNAs.
- Understanding protein-lncRNA interactions is critical for advancing research in molecular biology and disease mechanisms.
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01:20Immunoprecipitation
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
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02:39lncRNA - Long Non-coding RNAs
In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA (lncRNA)...
02:36Chromatin Immunoprecipitation- ChIP
Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
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ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
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