Cellular Membrane RNA Activities

University Of Colorado, Boulder CO

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Abstract

This work intends to survey specific RNAs associated with isolated membrane systems of cultured human cells. These RNAs are identified with specific sequences by array hybridization and massed sequencing. Identihed individual membrane-associated RNAs are subsequently screened for binding to purified phospholipid bilayers with the composition of human membranes, in the presence and absence of proteins. Residence on cellular membran es in vivo for RNAs with these biochemical properties is confirmed using FRET between RNA-complementary fluorescent oligonucleotides and membrane dyes introduced into living cells. The intended net effect ofthese three protocols is to detect a subset of membrane-resident RNAs in human cells that have bilayer affinity built into their covalent structures,