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Development and Optimization of MicroRNA against Relaxin‐3
Author(s) -
Callander Gabrielle E.,
Thomas Walter G.,
Bathgate Ross A. D.
Publication year - 2009
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2008.03816.x
Subject(s) - relaxin , microrna , gene knockdown , rna interference , in vivo , biology , microbiology and biotechnology , transfection , function (biology) , rna , bioinformatics , gene , endocrinology , genetics , hormone
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.