Development and optimization of microRNA against relaxin-3.
Recent in vivo studies suggest a role for relaxin-3 in feeding and stress. To further elucidate the function of relaxin-3 in the central nervous system, we have employed a complementary approach, based on RNA interference, to modulate relaxin-3 expression. We have designed, constructed, and characterized three microRNAs (miRNAs) targeting different regions of the relaxin-3 transcript. These were tested to determine the amount of miRNA required to achieve the greatest knockdown and for their ability to reduce the expression of relaxin-3 in transfected HEK293Tcells. All miRNA constructs significantly reduced relaxin-3-induced cAMP responses; however, miR499 was most effective. This should be a useful tool for in vitro studies and central targeting of relaxin-3 in vivo using viral delivery systems.
Department of Biochemistry and Molecular Biology, Howard Florey Institute, University of Melbourne, Parkville, Victoria, Australia.
Gabrielle E
GE
Callander
Walter G
WG
Thomas
Ross A D
RA
Bathgate
D002460:Cell Line; D005247:Feeding Behavior; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology; D012065:Relaxin / Q000235:genetics / Q000502:physiology*
D035683:MicroRNAs; C480238:RLN3 protein, human; D012065:Relaxin