Patent ID: 8846350
Filing Date: 2014-09-30
Classification: C12N,C12Q

Abstract:
1. A method for determining which microRNAs bind to a target mRNA comprising the steps of (a) creating a 3′UTR_RNA from the target mRNA by creating a double-stranded 3′UTR from the target mRNA by reverse transcription of the mRNA to synthesize cDNA and polymerase chain reaction (PCR) of the cDNA; and synthesizing 3′UTR_RNA from the double stranded 3′UTR PCR product using in vitro transcription; (b) creating a bait sequence from the target mRNA, where the bait sequence comprises a biotinylated cRNA of a 3′ untranslated region (UTR) that binds to a binding agent and where the bait sequence comprises the 3′UTR_RNA; (c) adding a mixture of microRNAs to the bait sequence; (d) separating the microRNAs that bind to the bait sequence from those microRNAs that do not bind; and (e) identifying the microRNAs that bind to the bait sequence, wherein the microRNAs identified are those that bind to the target mRNA.