title: Live Cell Genomics: RNA Exon-Specific RNA-Binding Protein Isolation.
abstract: RNA-binding proteins (RBPs) are essential regulatory proteins that control all modes of RNA processing and regulation. New experimental approaches to isolate these indispensable proteins under in vivo conditions are needed to advance the field of RBP biology. Historically, in vitro biochemical approaches to isolate RBP complexes have been useful and productive, but biological relevance of the identified RBP complexes can be imprecise or erroneous. Here we review an inventive experimental to isolate RBPs under the in vivo conditions. The method is called peptide nucleic acid (PNA)-assisted identification of RBP (PAIR) technology and it uses cell-penetrating peptides (CPPs) to deliver photo-activatible RBP-capture molecule to the cytoplasm of the live cells. The PAIR methodology provides two significant advantages over the most commonly used approaches: (1) it overcomes the in vitro limitation of standard biochemical approaches and (2) the PAIR RBP-capture molecule is highly selective and adaptable which allows investigators to isolate exon-specific RBP complexes. Most importantly, the in vivo capture conditions and selectivity of the RBP-capture molecule yield biologically accurate and relevant RBP data.
authors.affiliation: Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, 19104, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Bell
authors.forename: James
authors.initials: J
authors.lastname: Eberwine
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002470:Cell Survival; D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005091:Exons; D023281:Genomics / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D057846:Cell-Penetrating Peptides; D020135:Peptide Nucleic Acids; D016601:RNA-Binding Proteins; D012313:RNA