In vivo siRNA delivery to the mouse hypothalamus shows a role of the co-chaperone XAP2 in regulating TRH transcription.
RNA interference mediated by small interfering RNAs (siRNAs) is a powerful tool for evaluating gene function in vivo. In particular it should be able to provide tissue-specific and developmental stage-specific knockdown of target genes in physiological contexts. However, there are few demonstrations of its use on neuronal specific genes in vivo. We recently developed a cationic lipid-based approach to study gene function in a neuronal context. In particular, we applied it to study how the novel partner for TRbeta1, hepatitis virus B X-associated protein 2 (XAP2), a protein first identified as a co-chaperone protein, affects T3-transcriptional repression of the hypothalamic gene, TRH. The cationic lipid-based technique used, JetSI/DOPE, was previously shown to efficiently knockdown reporter gene mRNA in vivo. Using JetSI/DOPE to vectorize siRNA against XAP2 mRNA, we show that XAP2 is needed specifically for TRbeta1-mediated (but not TRbeta2) activation of hypothalamic TRH transcription. Thus, this cationic lipid-based siRNA strategy can effectively be used to reveal fine, tissue-specific and isoform-specific effects on neuronal gene transcription in vivo.
Decherf
Stéphanie
S
Laboratoire d'Evolution des Régulations Endocriniennes, Muséum National d'Histoire, Naturelle, Paris, France.
Hassani
Zahra
Z
Demeneix
Barbara A
BA
D000818:Animals; D000831:Animals, Newborn; D004210:Dissection; D005260:Female; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism*; D007267:Injections; D047908:Intracellular Signaling Peptides and Proteins; D008156:Luciferases / Q000378:metabolism; D008297:Male; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000494:pharmacology*; D013238:Stereotaxic Techniques; D013973:Thyrotropin-Releasing Hormone / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects*; D014284:Triiodothyronine / Q000494:pharmacology
D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D011506:Proteins; D034741:RNA, Small Interfering; C103265:aryl hydrocarbon receptor-interacting protein; D014284:Triiodothyronine; D013973:Thyrotropin-Releasing Hormone; D008156:Luciferases