Dark Ion Channel Overexpression Cell Lines

About this resource As part of the IDG Mission of providing resources for scientific community to enable study of the dark genes, we generated cell lines that over-express dark ion channels. These cell lines provide invaluable resource for biochemical, molecular and biophysical studies of ion channels.

How this resource was generated? cDNAs containing dark ion channels with V5 tag on the C-terminus in pLX304 vector were used to make lentivirus used to transduce early passage HEK293T cells for stable cell line generation. Cells were selected with Blasticidin for 10 days, expanded and then stained with V5 antibody to determine ion channel expression/localization. Images for all cells that survived the selection are shown below (regardless of the V5 status). These cells can be cultured in regular DMEM or RPMI media with 10% serum. Based on the presented imaging data, some cell lines express the channel well, while others do not appear to express. Although this is typical, the Ion Channel IDG is working on projects that will reveal factors important for ion channel expression and welcomes collaboration.

Where this resource can be requested These cells are available from UCSF Cell Engineering core (https://cgec.ucsf.edu/cell-culture-and-banking-services) upon request. If you have commercial interests, please include this in your email query.