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In Vitro Analysis of SERCA2 Gene Regulation in Hypertrophic Cardiomyocytes and Increasing Transfection Efficiency by Gene‐Gun Biolistics a
Author(s) -
EIZEMA KARIN,
HEUGTEN HAN A.A.,
BEZSTAROSTI KAREL,
SETTEN MARGA C.,
LAMERS JOS M.J.
Publication year - 1999
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1999.tb09229.x
Subject(s) - transfection , in vitro , gene , gene gun , microbiology and biotechnology , chemistry , biology , genetics , plasmid , dna vaccination
A bstract : The transcriptional downregulation of the SERCA2 gene is studied using neonatal rat cardiomyocytes stimulated with endothelin‐1 to induce hypertrophy. Liposome‐based transfection of cells with a 1.9 kb SERCA2 promoter fragment directed expression of a reporter gene identical to the down‐regulation of genomic SERCA2 expression by endothelin‐1. Results of a new gene gun technology for transient transfection of cardiomyocytes with a RSV‐β‐galactosidase construct are reported. This new method for propelling DNA‐coated gold beads into cardiomyocytes is extremely suitable for directly testing promoter/reporter gene DNA constructs since the transfection efficiency (approximately 10%) appears to be higher than traditional transfection methods.