siRNA-mediated Silencing of Peroxisomal Genes in Mammalian Cells
Methods in Molecular Biology
Humana Press (Springer Imprint)
Reason for embargo
RNAi technologies are a valuable tool in the identification and investigation of proteins which are involved in peroxisome biogenesis and function. Small interfering RNA (siRNA) has developed into the most commonly used RNAi tool for the induction of transient, shortterm silencing of protein coding genes. Although siRNA can induce gene knockdown in a variety of mammalian cell lines, their utility is limited by efficient uptake of synthetic oligonucleotides into the cells. Here, we describe different transfection methods which have been successfully used by us to silence peroxisomal genes in a variety of cell lines, including primary human skin fibroblasts, which are usually difficult to transfect.
We would like to thank J. Costello for his support and F. S. Alkuraya and P. Freisinger for providing Mff-deficient skin fibroblasts. This work was supported by the Marie Curie Initial Training Network (ITN) action (FP7-2012-PERFUME-316723) and the Biotechnology and] Biological Sciences Research Council (BB/K006231/1; BB/N01541X/1).
This is the author accepted manuscript. The final version is available from Humana Press via the DOI in this record.
Vol. 1595, pp 69-79