Journal
MOLECULES
Volume 26, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/molecules26082270
Keywords
RNA– protein complex; proximity labeling; biotin ligase; ascorbate peroxidase; RNA-binding protein; subcellular transcriptomics
Funding
- International Max Planck Research School (IMPRS) From Molecules to Organisms
- German Research Society [DFG-RTG2364]
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The interaction between RNAs and proteins controls multiple cellular functions and specific knowledge is required to understand their interactions. Various techniques have been developed to study RNA-protein interactions including CLIP and RNA affinity purification. In vivo proximity labeling can be used to study RNA-protein interactions and analyze tagged molecules via mass spectrometry or RNA-Seq.
Multiple cellular functions are controlled by the interaction of RNAs and proteins. Together with the RNAs they control, RNA interacting proteins form RNA protein complexes, which are considered to serve as the true regulatory units for post-transcriptional gene expression. To understand how RNAs are modified, transported, and regulated therefore requires specific knowledge of their interaction partners. To this end, multiple techniques have been developed to characterize the interaction between RNAs and proteins. In this review, we briefly summarize the common methods to study RNA-protein interaction including crosslinking and immunoprecipitation (CLIP), and aptamer- or antisense oligonucleotide-based RNA affinity purification. Following this, we focus on in vivo proximity labeling to study RNA-protein interactions. In proximity labeling, a labeling enzyme like ascorbate peroxidase or biotin ligase is targeted to specific RNAs, RNA-binding proteins, or even cellular compartments and uses biotin to label the proteins and RNAs in its vicinity. The tagged molecules are then enriched and analyzed by mass spectrometry or RNA-Seq. We highlight the latest studies that exemplify the strength of this approach for the characterization of RNA protein complexes and distribution of RNAs in vivo.
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