Integration efficiency of a hybrid adenoretroviral vector.
The frequency with which the hybrid vector AdLTR-luc mediates genomic integration [Nat. Biotech. 18 (2000) 176-180] is unknown. To address this, we constructed AdLTR-red, using the AdLTR-luc backbone and the enhanced red fluorescence protein cDNA. Kinetic studies showed that AdLTR-red and a conventional adenoviral vector, AdCMV-red, entered and transduced epithelial cells comparably. AdLTR-red integration frequency in vitro, i.e., the percentage of red clones after 10-12 doubling times from limiting dilutions, was 8.0% (36/450; at 67 particles/cell). With AdCMV-red, 0/549 clones were integration-positive. Seven days after AdLTR-luc or AdCMV-luc (10(11) particles) femoral vein administration to adult rats splenocytes were prepared, stimulated with concanavalin A, and examined by FISH. AdLTR-luc integration occurred in 5-11% of mitotic rat splenocytes, while no unequivocal integration was found with AdCMV-luc. These data provide the first quantitative evidence of the frequency for genomic integration with this hybrid vector.
Zheng
Changyu
C
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bldg. 10, Room 1N113, MSC 1190, Bethesda, MD 20892-1190, USA.
Wang
Jianghua
J
Baum
Bruce J
BJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D006824:Hybridization, Genetic; D008156:Luciferases / Q000235:genetics; D051381:Rats; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*; D016662:Virus Integration / Q000235:genetics*
D008156:Luciferases