内质网
基因沉默
细胞生物学
互补
小干扰RNA
生物
生物发生
细胞
转运蛋白
基因
转染
遗传学
表型
作者
Johanna Dudek,Sven Lang,Stefan Schorr,Johannes Linxweiler,Markus Greiner,Richard Zimmermann
出处
期刊:Methods in molecular biology
日期:2013-01-01
卷期号:: 285-299
被引量:5
标识
DOI:10.1007/978-1-62703-487-6_18
摘要
The development of small-interfering RNA (siRNA)-mediated gene-silencing strategies has made it possible to study the transport of precursors of soluble and membrane proteins into the endoplasmic reticulum (ER) of human cells. In these approaches, a certain target gene is silenced in the cell type of choice, followed by analysis of the effect of this silencing on the biogenesis of a single or set of precursor polypeptide(s) in cell culture or in cell-free assays involving semi-permeabilized cells and in vitro translations systems. These approaches allow for functional analysis of components of the ER-resident protein transport machinery as well as the elucidation of their potential cell-type variations and regulatory mechanisms. The gene-silencing and subsequent plasmid-based complementation carries the additional benefit of facilitating analysis of the consequences of disease-linked mutations in ER transport components.
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