A polyacrylamide microbead-integrated chip for the large-scale   manufacture of ready-to-use esiRNA

doi:10.1039/c0lc00564a

CORC > 北京大学 > 工学院

	A polyacrylamide microbead-integrated chip for the large-scale manufacture of ready-to-use esiRNA
	Huang, Huang ; Chang, Qing ; Sun, Changhong ; Yin, Shenyi ; Li, Juan ; Xi, Jianzhong Jeff
刊名	芯片实验室
	2011
关键词	RNA INTERFERENCE CELL-DIVISION CANCER
DOI	10.1039/c0lc00564a
英文摘要	Endoribonuclease-prepared siRNAs (esiRNAs) have the advantages of cost effectiveness and lower off-target effects than chemically synthesized siRNA. However, the current manufacture of esiRNA is a complex process, requiring an expensive instrument and demanding skills to accomplish the transfer, purification, quantification and normalization of liquid samples. These performances significantly hamper the application of esiRNAs on a large-scale level. In this study, we present a polymer microbead-integrated chip capable of the large-scale manufacture of esiRNA in a convenient and robust manner. This chip is able to perform the amplification, transcription and enzymatic digestion of targets on polymer scaffold, thus simplifying the transfer and purification manipulation process. What is also noted, this chip can readily tailor and normalize the amount of esiRNA product by controlling the number of DNA probes and the cycle of the amplification reaction. Thus the esiRNA, also referred to as gel-esiRNA, can be immediately applied to loss-of-function study without any further treatment. The silencing specificity and efficiency of gel-esiRNAs were assessed on transcriptional, translational or cell functional levels. All data of real-time PCR, Western blot assay, or FACS clearly supported that the gel-esiRNA produced specific gene silencing as effectively as the one generated following the conventional approach. We believe that this approach would provide a more robust and cost-effective choice to manufacture esiRNAs, thus promising both more intensive and extensive applications of these heterogeneous RNA strands.; Biochemical Research Methods; Chemistry, Multidisciplinary; Nanoscience & Nanotechnology; SCI(E); PubMed; 1; ARTICLE; 6; 1036-1040; 11
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/194365]
专题	工学院
推荐引用方式 GB/T 7714	Huang, Huang,Chang, Qing,Sun, Changhong,et al. A polyacrylamide microbead-integrated chip for the large-scale manufacture of ready-to-use esiRNA[J]. 芯片实验室,2011.
APA	Huang, Huang,Chang, Qing,Sun, Changhong,Yin, Shenyi,Li, Juan,&Xi, Jianzhong Jeff.(2011).A polyacrylamide microbead-integrated chip for the large-scale manufacture of ready-to-use esiRNA.芯片实验室.
MLA	Huang, Huang,et al."A polyacrylamide microbead-integrated chip for the large-scale manufacture of ready-to-use esiRNA".芯片实验室 (2011).