Screen Status: Primary Repeat
Screen Overview: Genome screen using 384 well library
Screener: Marco Gerlinger (Swanton Lab).
Extension:3462, Rm 115
Library: Full Genome 384
Library Description: Full Dharmacon 267 plate genome library stamped into 384 well plates
Library Supplier: Dharmacon
Library Format: 67 plate(s) in 384 well format
Replicate Info: 3 replicates
Biological replicates
Publications: Genome-wide RNA interference analysis of renal carcinoma survival regulators identifies MCT4 as a Warburg effect metabolic target.. Gerlinger et al., 2012
Keywords: Everolimus, Kidney cancer, Viability
Cell Line: RCC4

Screen Method

Day 1: Transfection of RCC4, A498 and 786-O cell lines

Cells were transfected with genome siRNA library (Dharmacon) using Dharmafect3 (RCC4) or LipoRNAiMAXi (A498 and 786-O) and cultured for 1 day.

Day 2: Addition of Everolimus

Everolimus is added on top of the culture media. For control plates, fresh media only was added. The plates were then cultured for further 3 days.

Day 5 Plates are fixed with 80% EthOH and stained with DAPI

Plates were fixed with 80% Ethanol and stained with DAPI.

Readout

Acumen Explorer eX3 laser scanning cytometer was used to determine cell number per well.