Stable integration of human immunodeficiency virus-based retroviral vectors into the chromosomes of nondividing cells.
Human immunodeficiency virus type 1 (HIV-1)-based vectors are thought to be useful for gene transfer into nondividing cells. We examined whether HIV vectors can really integrate into the chromosomes of nondividing cells. CD4+HeLa cells arrested at the G2 or G1/S phase were incubated with the HIV vector pseudotyped with the HIV envelope. The transduction efficiency of the HIV vector in these nondividing cells was comparable to that in proliferating cells. Sequencing of the polymerase chain reaction-amplified fragments containing the junction sites showed that the HIV vector was stably integrated into the chromosomal DNA. It was also demonstrated that terminally differentiated human macrophages and nonproliferating NT neurons could be transduced by the HIV vector after adenovirus-mediated expression of CD4. These results suggest that the HIV vector may be useful not only for gene therapy of AIDS but also for a variety of gene therapy protocols targeting nondividing cells.
Miyake
K
K
Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo, Japan.
Suzuki
N
N
Matsuoka
H
H
Tohyama
T
T
Shimada
T
T
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015704:CD4 Antigens / Q000235:genetics; D019556:COS Cells; D002455:Cell Division; D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000821:virology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000821:virology; D008979:Moloney murine leukemia virus / Q000235:genetics; D009474:Neurons / Q000821:virology; D017422:Sequence Analysis, DNA; D018243:Teratocarcinoma; D014162:Transfection; D014407:Tumor Cells, Cultured; D016662:Virus Integration
D015704:CD4 Antigens