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High‐level expression of active HIV‐1 integrase from a synthetic gene in human cells
Author(s) -
Cherepanov Peter,
Pluymers Wim,
Claeys Anje,
Proost Paul,
Clercq Erik,
Debyser Zeger
Publication year - 2000
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.14.10.1389
Subject(s) - integrase , integrase inhibitor , hek 293 cells , biology , gene , microbiology and biotechnology , cell culture , viral vector , viral replication , virology , genetics , virus , recombinant dna , antiretroviral therapy , viral load
A synthetic gene encoding for HIV‐1 integrase was designed to circumvent the intrinsic instability and the repressor elements present in the wild‐type gene. High‐level expression of HIV‐1 integrase was obtained in various human cell lines independently of viral accessory proteins. A human 293T cell line was selected that stably expresses HIV‐1 integrase and has growth kinetics comparable to the parental cell line. The enzyme was localized in the nucleus and remained stably associated with the chromosomes during mitosis. Lentiviral vector particles carrying the inactivating D64V mutation in the integrase gene were capable of stably transducing 293T cells when complemented in the producer cells with integrase expressed from the synthetic gene. When the cell line that stably expresses integrase was infected with the defective viral particles, complementation of integrase activity was detected as well. Expression of active HIV‐1 integrase in human cells will facilitate the study of the interplay between host and viral factors during integration.—Cherepanov, P., Pluymers, W., Claeys, A., Proost, P., De Clercq, E., Debyser, Z. High‐level expression of active HIV‐1 integrase from a synthetic gene in human cells. FASEB J. 14, 1389–1399 (2000)