caa a22 4500 445292687 CHVBK 20180317142533.0 cr unu---uuuuu 170323e20101201xx s 000 0 eng 10.1007/s11626-010-9347-6 doi (NATIONALLICENCE)springer-10.1007/s11626-010-9347-6 Stable suppression of gene expression by short interfering RNAs targeted to promoter in a mouse embryonal carcinoma stem cell line [Elektronische Daten] [Fariba Esmaeili, Taravat Bamdad, Sorayya Ghasemi] RNA interference (RNAi) can induce gene silencing via two pathways: post-transcriptional gene silencing (PTGS) and transcriptional gene silencing (TGS). The mediators of gene inactivation in both pathways are 21-bp small interfering RNAs (siRNAs) generated from longer double-stranded RNA (dsRNA). PTGS involves siRNA-mediated targeting and degradation of mRNA. However, siRNAs induce TGS via DNA methylation at the targeted promoter. Synthetic siRNAs can induce loss of gene activity comparable to long dsRNA. The limitation of this method is that the transfected synthetic siRNA works for only a few days. In this study, we tested the RNAi response to siRNA (PTGS pathway) by using a plasmid containing an enhanced green fluorescent protein (eGFP) gene as a target as well as a plasmid creates siRNA transcript, in a form of a hairpin, against eGFP gene. To investigate TGS pathway via RNAi, we also used a plasmid creates hairpin siRNA transcript against pgk-1 promoter. The data presented here indicated long-lasting inhibition in expression of eGFP and puromycin genes, both under the control of the murine Pgk-1 promoter. However, Southern blot analysis showed no methylation in pgk-1 promoter. The Society for In Vitro Biology, 2010 DNA methylation nationallicence Gene silencing nationallicence Promoter nationallicence RNA interference nationallicence Small interfering RNA nationallicence Esmaeili Fariba Department of Biology, Faculty of Basic Sciences, Shahr-e-kord University, P.O. Box115, Shahr-e-kord, Iran aut Bamdad Taravat Department of Virology, School of Medical Sciences, Tarbiat Modarres University, P.O. Box14115-331, Tehran, Iran aut Ghasemi Sorayya Cellular & Molecular Research Center, Shahr-e-kord University of Medical Sciences, Shahr-e-kord, Iran aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/10(2010-12-01), 834-840 1071-2690 46:10<834 2010 46 11626 https://doi.org/10.1007/s11626-010-9347-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9347-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Esmaeili Fariba Department of Biology, Faculty of Basic Sciences, Shahr-e-kord University, P.O. Box115, Shahr-e-kord, Iran aut NATIONALLICENCE 700 1- Bamdad Taravat Department of Virology, School of Medical Sciences, Tarbiat Modarres University, P.O. Box14115-331, Tehran, Iran aut NATIONALLICENCE 700 1- Ghasemi Sorayya Cellular & Molecular Research Center, Shahr-e-kord University of Medical Sciences, Shahr-e-kord, Iran aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/10(2010-12-01), 834-840 1071-2690 46:10<834 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer