[{"id":374,"title":"Novel method to isolate and identify specific RNA/DNA binding proteins","sha":"aca025321ad0189d24927bdfeca289ef","user_id":null,"state":"published","body":"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.\r\n\r\n**_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.\r\n\r\nSimilarly, 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.\r\n","created_at":"2017-03-27T19:50:17.659Z","updated_at":"2017-04-15T15:47:28.504Z","zenodo_id":438357,"doi":"https://doi.org/10.5281/zenodo.438357","subject":null,"tags":["rna binding proteins","dna binding proteins","neurodegenration","cell asymmetry"],"vote_count":5,"view_count":1,"score":5.1,"deleted":false,"muted":false,"tweeted":false,"attachment_file_name":null,"attachment_content_type":null,"attachment_file_size":null,"attachment_updated_at":null}]