The recombinant human parvoviruses for gene therapy of hemoglobinopathies.
Towards a goal of using adeno-associated viruses (AAV), the human parvovirus, as the gene transfer vector for gene therapy of hemoglobinopathies, the human beta-globin (h beta G) cDNA was ligated downstream of the P40 promoter of AAV type 2 (AAV2) genome. Transfection via electroporation of the construct into human 293 cells (embryonal kidney cell line) resulted in expression of the cloned h beta G cDNA, as evidenced by the synthesis of transcripts hybridizable to h beta G probe. The transfection led to the recombinant genome to be excised out of the plasmid and replicate in the cell, followed by production of the recombinant AAV that harbors h beta G cDNA.
Dixit
M
M
Department of Biochemistry, Meharry Medical College, Nashville, TN 37208.
Tillery
M K
MK
Plonk
S G
SG
Ohi
S
S
D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015316:Genetic Therapy; D005914:Globins / Q000235:genetics*; D006453:Hemoglobinopathies / Q000628:therapy*; D006801:Humans; D010321:Parvoviridae / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
D005914:Globins