Intracytoplasmic gene delivery for in vitro transfection with cytoskeleton-specific immunoliposomes

A novel and highly efficient method of in vitro gene transfection has been developed. This method employs direct intracytoplasmic gene delivery into e mbryonic cardiocytes using neutral cytoskeletal-antigen specific immunolipo somes (CSIL). These immunoliposomes target cardiocytes specifically under r eversible hypoxic conditions. Two independent reporter genes, pGL2 and pSV- beta -galactosidase, were used! to verify CSIL-transfection (CSIL-fection). The efficiency of CSIL-fection with firefly luciferase pGL2 vector was 30 times greater than controls consisting of hypoxic cardiocytes treated with plain liposomes (PL) or normoxic cardiocytes treated with CSIL, PL or nake d DNA. CSIL-fection was also compared to cationic liposome transfection. Ne t cationic liposome transfection appeared to be more efficient than CSIL-fe ction for pGL2 vectors. However, a smaller number of viable cells was obser ved in the cationic liposome treated cultures than in the CSIL treated cult ures. Therefore, to determine whether more cells were transfected with cati onic liposomes or CSIL, pSV-beta -galactosidase vector was used. CSIL-fecti on with pSV-beta -galactosidase vector produced at least 40 times more tran sfected cells than those transfected with cationic liposomes. No transfecti on with pSV-beta -galactosidase vectors was obtained with IgG-liposome, PL or naked DNA treatments. Targeted enhanced efficiency of transfection by th is novel method could have practical therapeutic applications in the geneti c modification of cells ex vivo that could then be reimplanted into patient s for gene therapy. (C) 2001 Elsevier Science BY All rights reserved.