Transcriptomics

Dataset Information

5

MicroRNA-like off-target transcript regulation by siRNAs


ABSTRACT: siRNAs mediate sequence-specific gene silencing in cultured mammalian cells but also silence unintended transcripts. Many siRNA off-target transcripts match the guide-strand ‘‘seed region,’’ similar to the way microRNAs match their target sites. The extent to which this seed-matched, microRNA-like, off-target silencing affects the specificity of therapeutic siRNAs in vivo is currently unknown. Here, we compare microRNA-like off-target regulations in mouse liver in vivo with those seen in cell culture for a series of therapeutic candidate siRNAs targeting Apolipoprotein B (APOB). Each siRNA triggered regulation of consistent microRNA-like off-target transcripts in mouse livers and in cultured mouse liver tumor cells. In contrast, there was only random overlap between microRNA-like off-target transcripts from cultured human and mouse liver tumor cells. Therefore, siRNA therapeutics may trigger microRNA-like silencing of many unintended targets in vivo, and the potential toxicities caused by these off-target gene regulations cannot be accurately assessed in rodent models. Hepa1-6 mouse hepatoma cell line and HUH7 and PLC/PRF/5 human hepatoma cell lines were transfected in 6-well plates using Lipofectamine RNAiMAX and siRNA duplexes at a final concentration of 10 nM. For in vitro analysis, RNA was extracted at 6, 12, 24, and 48 h post-transfection. For in vivo studies, mouse livers were harvested 3 d following a single administration of APOB siRNA (3 mg/kg) formulated in lipid nanoparticles. For details, please see: J. Burchard, A.L. Jackson, V. Malkov, R.H.V. Needham, Y. Tan, S.R. Bartz, H. Dai, A.B. Sachs and P.S. Linsley. microRNA-like off-target transcript regulation by siRNAs is species specific. RNA 15 (2009)

ORGANISM(S): Musculus  

SUBMITTER: Julja Burchard  Peter S Linsley  Vladislav Malkov  Aimee L Jackson  Yejun Tan  Alan B Sachs  Hongyue Dai  Steven R Bartz  Rachel H Needham 

PROVIDER: E-GEOD-14073 | ArrayExpress | 2009-01-31

SECONDARY ACCESSION(S): GSE14073PRJNA110373

REPOSITORIES: GEO, ArrayExpress

altmetric image

Publications

MicroRNA-like off-target transcript regulation by siRNAs is species specific.

Burchard Julja J   Jackson Aimee L AL   Malkov Vladislav V   Needham Rachel H V RH   Tan Yejun Y   Bartz Steven R SR   Dai Hongyue H   Sachs Alan B AB   Linsley Peter S PS  

RNA (New York, N.Y.) 20090201 2


siRNAs mediate sequence-specific gene silencing in cultured mammalian cells but also silence unintended transcripts. Many siRNA off-target transcripts match the guide-strand "seed region," similar to the way microRNAs match their target sites. The extent to which this seed-matched, microRNA-like, off-target silencing affects the specificity of therapeutic siRNAs in vivo is currently unknown. Here, we compare microRNA-like off-target regulations in mouse liver in vivo with those seen in cell cult  ...[more]

Similar Datasets

2009-02-01 | GSE14073 | GEO
2006-09-12 | E-GEOD-5814 | ArrayExpress
2010-11-22 | E-GEOD-23088 | ArrayExpress
2010-11-22 | GSE23088 | GEO
| GSE22209 | GEO
2006-09-07 | GSE5769 | GEO
| GSE68154 | GEO
2016-10-12 | PXD004141 | Pride
2007-04-10 | GSE6807 | GEO
| GSE78084 | GEO