About this resource
As part of the IDG Mission of providing resources for scientific community to enable study of the dark genes, we generated dark ion channels expression constructs in lentiviral backbone. These constructs allow stable lentiviral transduction and selection for the ion channel of interest and were used to generate cell lines described under “Cells” resource tab.
How this recourse was generated?
Plasmids containing coding sequences for dark ion channel genes (longest splice isoform whenever possible) were obtained in various formats from the hORFeome, and other commercial sources. For plasmids from hORFeome collection, ORFs were already in pLX304 vector, which were used directly for lentivirus production. For plasmids in non-pLX304 backbone ion channel ORFs were cloned into pLX304 vector using Gateway and In-Fusion strategies. Reaction mixtures for all pipelines were transformed into Stbl3 bacteria and Plasmid DNA was isolated for validation of the ion channel coding part by Sanger sequencing. All available cDNAs in pLX304 vector that were sequence verified by Sanger are listed below.
Where this resource can be requested
These constructs are available at McManus lab UCSF upon request. We are currently in the process of depositing these constructs at Addgene as a collection. If you have commercial interests, please include this in your email query.