RNA-seq of total royal jelly and MRJP-3-bound RNA

MRJP-3 is an RNA binding protein that binds both double- and single-stranded RNA in vitro. To test whether i) MRJP-3 binds RNA in its natural environment, the royal jelly (RJ) and ii) to characterize its RJ RNA partners; a pull down followed by RNA-seq system was developed. MRJP-3 bound RNA was compared to total RJ RNA extracted from the same hive. To establish the system, we first incubated biotinylated MRJP-3, or biotinylated BSA in RJ and pulled the proteins out with strepavidin coated magnetic beads followed by RNA extraction and bioanalyzer profiling. MRJP-3 pull out was enriched for RNA compare to the BSA and just beads controls. The RNA population pulled down with MRJP-3 has similar profile to the total RJ RNA, further demonstrating the MRJP-3 has no affinity to a certain RNA species.