Targeting of A701G nucleotide at the human ATP1A1 locus using a RNA/DNA chimera.
The single base substitution mediated by chimeric RNA/DNA oligonucleotide is a new promising approach of gene therapy for single base mutation diseases. We exploited this approach to render HeLa cells resistant to ouabain by introducing a single base substitution in the alpha 1 subunit of the NA+/K+ ATPase human gene. The chimeric oligonucleotide was administered to HeLa cells by electroporation and the frequency of ouabain resistant cells determined. The results showed that the chimeric RNA/DNA oligonucleotide failed to enhance the frequency of ouabain resistant cells supporting the controversy about the conflicting results of the technique.
Cervelli
Tiziana
T
Laboratorio di Terapia Genica e Molecolare, Istituto di Fisiologia Clinica, Area della Ricerca del CNR, via G. Moruzzi, 1, 56127 Pisa, Italy.
Lombardi
Grazia
G
Citti
Lorenzo
L
Galli
Alvaro
A
Locci
Maria Teresa
MT
Rainaldi
Giuseppe
G
D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D018274:Electroporation; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000235:genetics*; D017354:Point Mutation / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics*
D009711:Nucleotides; D012313:RNA; D004247:DNA; D000254:Sodium-Potassium-Exchanging ATPase