Nucleofection and RNAi
Gene silencing by RNA interference (RNAi) emerged as a powerful technology for assessing gene function in mammalian cells, and has become a valuable tool in functional genomics, target discovery and validation.
The two most critical factors determining the effectiveness of RNAi experiments are the ability of the siRNA sequence to specifically silence the target mRNA, and the efficiency with which siRNA (or an shRNA-expressing vector) can be transfected into the cells of interest.
With over 400 RNAi related
publications, nucleofection has proven to be the delivery method of choice for any RNAi substrate. The unique versatility of the technology allows for a wide range of research applications from basic research studies such as analyzing the mechanisms of microRNA, to functional studies via RNAi mediated gene knockdown. In addition, the 96-well Shuttle now offers the possibility to perform RNAi library screenings using siRNA or shRNA in more relevant cell types such as primary cells (e.g. neurons) and difficult-to-transfect cell lines (e.g. Jurkats).
Efficient non-viral delivery
- RNAi experiments in difficult-to-transfect cell types, e.g. Jurkat or primary neurons
- High transfection efficiencies for different substrates, e.g. RNA oligonucleotides (
siRNA, synthetic
miRNA, miRNA inhibitors) and
shRNA- or miRNA-expressing vectors
Optimized protocols
- Ready-to-use protocols for
primary cells and
cell lines - Same protocol for different substrates allows easy switch between substrates and co-transfection of different substrates
- High-throughput
RNAi screenings in primary cells and difficult-to-transfect cell lines
Non reagent-based transfection
- High reproducibility
- No reagent-induced cytotoxicity
- No lipid-induced induction of interferon responses1
- No lipid-induced off-target effects2
1) Marques JT & Williams RG (2005) Nat Biotechnol 23(11):1399-405
2) Fedorov Y et al. (2005) Nat Methods 2(4):241

