Using DT40 to study clathrin function.
Clathrin-coated pits and vesicles play a major role in eukaryotic membrane trafficking pathways. We have used the DT40 system to delete endogenous clathrin genes selectively from DT40 and replace them with clathrin under the control of a tetracycline-regulatable promoter. This enabled clathrin expression to be manipulated, and the functional consequences of clathrin depletion to be studied in a stable vertebrate context. Here we describe the background to the work on clathrin, our practical experience with using the tetracycline-regulatable expression system in DT40 and some novel insights into membrane trafficking pathways obtained using the cell-lines generated during the course of this work.
Wettey
Frank R
FR
Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, UK.
Jackson
Antony P
AP
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology*; D002460:Cell Line; D002645:Chickens; D002966:Clathrin / Q000502:physiology*; D004705:Endocytosis; D006056:Golgi Apparatus / Q000378:metabolism; D008561:Membrane Fusion
D002966:Clathrin