Elimination of the truncated message from the herpes simplex virus thymidine kinase suicide gene.
Introduction of the Herpes simplex virus thymidine kinase (HSV-tk) gene into target cells renders them susceptible to killing by ganciclovir (GCV). We are studying the use of HSV-tk-transduced T lymphocytes in the context of hematopoietic stem cell transplantation. We have previously shown, in vitro and in vivo, the occurrence of transduced cells resistant to GCV due to a deletion within HSV-tk. This deletion, a consequence of the presence of cryptic splice donor and acceptor sites, originates in the retroviral producer cell. Here we adopt two different methods that introduce third-base degenerate changes at the cryptic splice sites and so prevent splicing. Consequently, the HSV-tk protein is unaltered and the sensitivity of the target cells to GCV is preserved. The use of this mutated HSV-tk should reduce the likelihood of the development of resistant genetically modified cells during clinical trials.
Chalmers
D
D
Laboratoire de Thérapeutique Immuno-Moléculaire, INSERM-E0119, UPRES-EA 2284 Etablissement Français du Sang, Bourgogne/Franche-Comté, 1 Boulevard A. Fleming, Besançon, 25020, France. david.chalmers@efs.sante.fr
Ferrand
C
C
Apperley
J F
JF
Melo
J V
JV
Ebeling
S
S
Newton
I
I
Duperrier
A
A
Hagenbeek
A
A
Garrett
E
E
Tiberghien
P
P
Garin
M
M
D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D015774:Ganciclovir / Q000494:pharmacology; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic
D000998:Antiviral Agents; D013937:Thymidine Kinase; D015774:Ganciclovir