By Venu Gurram

Owing to their role in establishing cell asymmetry and their role in pathological conditions such as neurodegenerative diseases, it is very important to focus on eukaryotic RNA-binding proteins(RBPs) binding to a specific transcript(s). Although the biotin-conjugated method is currently in use to isolate RBPs, it has certain limitations such as background interference. Here, I am proposing a new idea which is simple, yet effective.

To isolate RBPs binding to specific RNA transcript, we can introduce a prokaryotic RBP binding site in the transcript of our interest while simultaneously co-expressing the corresponding prokaryotic RBP. Followed by RNA-Immunoprecipitation and mass spectrometry, we can identify RBPs binding to a specific transcript. This method, which is first of its kind, could have many advantages like identifying RBPs of prokaryotic-site linked endogenous transcripts(using genetically modified animals) while still retaining the native secondary structure of RNA.

Similarly, to isolate DNA binding proteins that bind to a specific DNA sequence(Promoter), we can introduce a prokaryotic DNA sequence in the DNA segment of our interest and co-express the corresponding prokaryotic DNA binding protein. This can later be followed by Chromatin Immunoprecipitation and mass spectrometry.


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Venu Gurram



Published: 27 Mar, 2017

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