A Magnetic Bead-Integrated Chip for the Large Scale Manufacture of   Normalized esiRNAs

doi:10.1371/journal.pone.0039419

CORC > 北京大学 > 工学院

	A Magnetic Bead-Integrated Chip for the Large Scale Manufacture of Normalized esiRNAs
	Wang, Zhao ; Huang, Huang ; Zhang, Hanshuo ; Sun, Changhong ; Hao, Yang ; Yang, Junyu ; Fan, Yu ; Xi, Jianzhong Jeff
刊名	plos one
	2012
关键词	ENDORIBONUCLEASE-PREPARED SIRNA RNA INTERFERENCE CELL-DIVISION SCREEN
DOI	10.1371/journal.pone.0039419
英文摘要	The chemically-synthesized siRNA duplex has become a powerful and widely used tool for RNAi loss-of-function studies, but suffers from a high off-target effect problem. Recently, endoribonulease-prepared siRNA (esiRNA) has been shown to be an attractive alternative due to its lower off-target effect and cost effectiveness. However, the current manufacturing method for esiRNA is complicated, mainly in regards to purification and normalization on a large-scale level. In this study, we present a magnetic bead-integrated chip that can immobilize amplification or transcription products on beads and accomplish transcription, digestion, normalization and purification in a robust and convenient manner. This chip is equipped to manufacture ready-to-use esiRNAs on a large-scale level. Silencing specificity and efficiency of these esiRNAs were validated at the transcriptional, translational and functional levels. Manufacture of several normalized esiRNAs in a single well, including those silencing PARP1 and BRCA1, was successfully achieved, and the esiRNAs were subsequently utilized to effectively investigate their synergistic effect on cell viability. A small esiRNA library targeting 68 tyrosine kinase genes was constructed for a loss-of-function study, and four genes were identified in regulating the migration capability of Hela cells. We believe that this approach provides a more robust and cost-effective choice for manufacturing esiRNAs than current approaches, and therefore these heterogeneous RNA strands may have utility in most intensive and extensive applications.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000305825800032&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Multidisciplinary Sciences; SCI(E); PubMed; 1; ARTICLE; 6; e39419; 7
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/153952]
专题	工学院
推荐引用方式 GB/T 7714	Wang, Zhao,Huang, Huang,Zhang, Hanshuo,et al. A Magnetic Bead-Integrated Chip for the Large Scale Manufacture of Normalized esiRNAs[J]. plos one,2012.
APA	Wang, Zhao.,Huang, Huang.,Zhang, Hanshuo.,Sun, Changhong.,Hao, Yang.,...&Xi, Jianzhong Jeff.(2012).A Magnetic Bead-Integrated Chip for the Large Scale Manufacture of Normalized esiRNAs.plos one.
MLA	Wang, Zhao,et al."A Magnetic Bead-Integrated Chip for the Large Scale Manufacture of Normalized esiRNAs".plos one (2012).