RNAi Screening for Target Identification

Nucleofection of siRNA Libraries

High-throughput transfection of siRNA libraries has become an indispensable tool in target identification and validation. However, meaningful siRNA screening results strongly depend on efficient delivery of siRNA into the selected cell type. Most screenings have so far been limited to easy-to-transfect adherent cell lines. The  Nucleofector 96-well Shuttle System now enables these approaches in more relevant cell types - primary and hard-to-transfect cells - with 

• Up to 99% transfection efficiency for siRNA
• High reproducibility
• Low cytotoxicity
• Ease to automate
• Circumvention of lipid-mediated interferon responses

siRNA Library Screens in Hard-to-Transfect HUVEC and Jurkat Cells

In collaboration with Thermo Scientific (Dharmacon Products), amaxa performed two different siRNA library screens using the 96-well Shuttle:

1. Primary human umbilical vein endothelial cells (HUVEC) were screened with a combined protein kinase and a cell-cycle library to identify targets that affect cell viability.
2. Jurkat T cells were screened with an apoptosis library to identify genes that regulate FAS-mediated cell death (see figure below).

The setup and results of both screens are summarized in our joint  Application Note.