mRNA transfection of dendritic cells: synergistic effect of ARCA mRNA capping with Poly(A) chains in cis and in trans for a high protein expression level

M. Mockey (1) , C. Goncalves (1) , F.P. Dupuy (2) , F.M. Lemoine (2) , C. Pichon (1) , P. Midoux (1)

1 CBM - Centre de biophysique moléculaire
2 BTPI - Biologie et thérapeutique des pathologies immunitaires

The occurrence of translation mechanism in the cytosol offers advantages to mRNA transfer over DNA-based transfection in non-dividing cells. Here, we sought to optimize mRNA constructs allowing a high level of protein upon lipofection. We found that luciferase into mouse dendritic cells (JAWSII cells) was not, vert, similar20-fold higher when the luciferase mRNA was capped with 3′-O-methyl-m75′Gppp5′G (anti-reverse cap analogue; ARCA) than with m75′Gppp5′G (CAP). Adding a Poly(A) tail of 100 instead of 64 adenosines in cis increased by not, vert, similar35-fold more the protein level. Finally, ARCA-Luc-mRNA-A100 construct was 700-fold better efficient than the CAP-Luc-mRNA-A64 one. Moreover, co-transfection with free Poly(A) chains in trans enhanced by 100% the luciferase level. The efficiency of ARCA-mRNA-A100 construct was validated in immature and mature human CD34-derived dendritic cells. Such mRNA construct was also successful to obtain high level of MART-1 tumor antigen.

Mots clés: mRNA transfection; Dendritic cells; Capping; Poly(A) chains; Cap; 3′UTR; 5′UTR; Lipofection

M. Mockey, C. Goncalves, F.P. Dupuy, F.M. Lemoine, C. Pichon, et al.. mRNA transfection of dendritic cells: synergistic effect of ARCA mRNA capping with Poly(A) chains in cis and in trans for a high protein expression level. Biochemical and Biophysical Research Communications, 2006, 340, pp.1062-1068. ⟨10.1016/j.bbrc.2005.12.105⟩. ⟨hal-00088921⟩