Elongation of peroxisomes as an indicator for efficient dynamin-like protein 1 knock down in mammalian cells.
RNA interference has become a valuable tool to identify and investigate proteins involved in the formation of peroxisomes. We demonstrate that the elongation of peroxisomes serves as an excellent indicator for efficient knock down of dynamin-like protein 1 (DLP1) in mammalian cells. We took advantage of the silencing-dependent morphological changes of peroxisomes to compare different transfection methods and show that a single transfection of DLP1 siRNA by electroporation is sufficient to effectively silence DLP1. We present a fast, easy, and convenient protocol for efficient gene silencing in a large number of cells, which can be used for quantitative and biochemical studies.
Boll
Antje
A
Department of Cell Biology and Cell Pathology, University of Marburg, Marburg, Germany.
Schrader
Michael
M
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D034281:Dynamins; D018274:Electroporation; D005455:Fluorescent Antibody Technique; D020558:GTP Phosphohydrolases / Q000235:genetics*; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D024101:Mitochondrial Proteins / Q000235:genetics*; D020675:Peroxisomes / Q000648:ultrastructure*; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins