5, (epub ahead of print) e10557.
Discovery and validation of sub-threshold genome-wide association study loci using epigenomic signatures.
Wang, X., Tucker, N.R., Rizki, G., Mills, R., Krijger, P.H.L., de Wit, E., Subramanian, V., Bartell, E., Nguyen, X.-X., Ye, J., Leyton-Mange, J., Dolmatova, E.V., van der Harst, P., de Laat, W., Elinor, P.T., Newton-Cheh, C., Milan, D.J., Kellis, M. and Boyer, L.A.
Notes: Putative enhancer elements were cloned into the pGL4.23 [luc2/minP] and transfected into human iCMs (Cellular Dynamics) along with a Renilla luciferase control. The iCMs were plated at 2 × 103 per well of a 96-well plate and cultured until all cells were electrically connected and beat simultaneously (~7 days post plating). Beating cells were transfected with the two reporters using the ViaFect™ Transfection Reagent at a 2:1 Viafect:DNA ratio (further detail provided in the paper). The reporter activities were measured 24 hours post-transfection with the Dual-Luciferase® Reporter Assay System. Transfections using a GFP-expressing vector provided a visual estimation of the transfection efficiency and 65-70% of the iCMs were GFP-positive 24 hrs post-transfection. (4671)
Expand Full Notes »