Reagent 007 for chemical transfection
Reagent 007 is a novel peptide-based vehicle for efficient delivery of nucleic acids that could be used for transfection of mammalian and insect cells.
The transfection efficiency in CHO cell lines in serum-free conditions is between 70-90%.
R007 transfection efficiencies and post-transfection viability of Icosagen’s proprietary protein production CHO cultures growing as suspension cells in chemically defined serum-free medium. (A) Transfection efficiencies measured by FACS analysis 24 h after transfection (B) Viabilities of the 20 ml scale cultures transfected with 10 μg of scFv-Fc antibody expression vector using Reagent 007. Culture viability was measured before transfection as well as 24, 48 and 72 hours post-transfection using Trypan blue exclusion. Picture modified from Karro et al., Methods Mol Biol. 2015; 1324:435-45
A real highlight for Reagent 007 is its suitability for the production of pseudotyped virus-like particles (VLPs) and excellent cell recovery. All other chemical transfection reagents we tested cause mislocalization of the membrane protein, which results in reduced production of the pseudotyped VLPs.
Features and advantages of Reagent 007:
- Non-lipid formulation
- Efficient transfection and very low toxicity
- Universal for delivery of diverse biomolecules
- Convenient and rapid procedure (complex formation time 5 min)
- Suitable for transfection of various eukaryotic cell lines in serum-free conditions: CHO, 293 and Sf9
- Suitable for production of virus-like particles (VLPs)
- Stable up to 1 year at -20°C
Reagent 007, 10 mg (powder)
Reagent 007 transfection protocol for suspension mammalian cells in serum-free conditions
- Arukuusk, P et al, New generation of efficient peptide-based vectors, NickFects, for the delivery of nucleic acid. Suhorutsenko J, Oskolkov N, Arukuusk P, Kurrikoff K, Eriste E, Copolovici DM, Langel U. Biochimica et Biophysica Acta, (2013) 1365-1373.
During the period 12 December 2013 to 15 April 2014, Icosagen AS carried out the project “Product and service design for marketing a brand new transfection reagent”, with support and co-financing from the European Union/European Social Fond. The project received support from the Enterprise Estonia program “Development of knowledge and skills”.