Acoustic Liquid Handling for Rapid siRNA Transfection Optimization.
Gene knockdown by small interfering RNA (siRNA) has been used extensively to investigate the function of genes in targeted and genome-wide studies. One of the primary challenges of siRNA studies of any scale is to achieve sufficient gene knockdown to produce the biological changes that lead to measurable phenotypes. Reverse, lipid-based transfection efficiency minimally requires the optimization of the following parameters: cell number, knockdown duration, siRNA oligonucleotide concentration, type/brand of transfection lipid, and transfection lipid concentration. In this study, we describe a methodology to utilize the flexibility and low-volume range of the Echo acoustic liquid handler to rapidly screen a matrix of transfection conditions. The matrix includes six different transfection lipids from three separate vendors across a broad range of concentrations. Our results validate acoustic liquid transfer for the delivery of siRNAs and transfection reagents. Finally, this methodology is applied to rapidly optimize transfection conditions across many tissue culture cell lines derived from various originating tissues.
Takeda Pharmaceuticals International Co., Cambridge, MA, USA.
Andrew S
AS
Xiao
Takeda Pharmaceuticals International Co., Cambridge, MA, USA.
Eric S
ES
Lightcap
Takeda Pharmaceuticals International Co., Cambridge, MA, USA david.bouck@takeda.com.
David C
DC
Bouck
D045744:Cell Line, Tumor; D002470:Cell Survival; D055785:Gene Knockdown Techniques; D018014:Gene Transfer Techniques; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D015203:Reproducibility of Results; D014162:Transfection / Q000379:methods*
D034741:RNA, Small Interfering
acoustic liquid handling; siRNA transfection