CAP-MAP: Cap Analysis Protocol with Minimal Analyte Processing, a rapid and sensitive approach to analysing mRNA cap structures

Abstract

Eukaryotic messenger RNA (mRNA) is modified by the addition of an inverted guanosine cap to the triphosphate at the 5 end. The cap guanosine and initial transcribed nucleotides are further methylated by a series of cap methyltransferases to generate the mature cap structures which protect RNA from degradation and recruit proteins involved in RNA processing and translation. Research demonstrating that the cap methyltransferases are regulated has generated interest in determining the methylation status of the mRNA cap structures present in cells. Here we present CAP-MAP: Cap Analysis Protocol with Minimal Analyte Processing, a rapid and sensitive method for detecting cap structures present in mRNA isolated from tissues or cell cultures.