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MS2 tagging

MS2 tagging is a technique based upon the natural interaction of the MS2 bacteriophage coat protein with a stem-loop structure from the phage genome, which is used for biochemical purification of RNA-protein complexes and partnered to GFP for detection of RNA in living cells. More recently, the technique has been used to monitor the appearance of RNA in living cells, at the site of transcription, or simply by observing the changes in RNA number in the cytoplasm. This has revealed that transcription of both prokaryotic and eukaryotic genes occurs in a discontinuous fashion with bursts of transcription separated by irregular intervals.

Procedure
Start with single-stranded RNA, and create a pattern of stem-loop structures by adding copies of the MS2 RNA-binding sequences to a noncoding region. The MS2 protein must be fused with GFP and bonded to an mRNA, a complex that contains the MS2’s RNA-binding sequence copies. MS2 biotin-tagged RNA affinity purification (MS2-BioTRAP) is one in vivo method of identifying protein-RNA interactions. Both the RNA that tagged with MS2 and the MS2 protein tag were expressed, and then, the affinity interaction was used to help the process of identifying protein-RNA interactions. == Advantages and Disadvantages ==
Advantages and Disadvantages
Advantages: The MS2-BioTRAP method is fast, flexible, and easy to set up; it scales well and allows the study of the physiological conditions of the protein-RNA interactions. Also, by using MS2 as an affinity tag to purify a protein in E. coli bacteria, scientists expressed MS2-MBP, which is an MS2 coat protein carrying mutations fused with maltose-binding proteins. The mutations prevented oligomerization. == References ==
Source: Wikipedia ↗
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