Delivery of the Cre Recombinase by a Self-Deleting Lentiviral Vector: Efficient Gene Targeting in vivo

The Cre recombinase (Cre) from bacteriophage P1 is an important tool for genetic engineering in mammalian cells. We constructed lentiviral vectors that efficiently deliver Cre in vitro and in vivo. Surprisingly, we found a significant reduction in proliferation and an accumulation in the G2/M phase...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2001-09, Vol.98 (20), p.11450-11455
Main Authors: Pfeifer, Alexander, Brandon, Eugene P., Kootstra, Neeltje, Gage, Fred H., Verma, Inder M.
Format: Article
Language:English
Subjects:
Animals
B lymphocytes
Biological Sciences
Bone marrow cells
Bone Marrow Cells - enzymology
Brain - enzymology
Cell Cycle
Cell Division
Cell growth
Cell Line
Cell Survival
Cells
Cells, Cultured
Cercopithecus aethiops
COS cells
Experiments
Gene Expression Regulation, Enzymologic
Genes, Reporter
Genetic engineering
Genetic Vectors
Genetics
Hepatocytes
Infections
Integrases - genetics
Lentivirus
Lentivirus - enzymology
Mammals
Mice
Mice, Inbred Strains
Stem cells
Viral Proteins - genetics
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Summary:The Cre recombinase (Cre) from bacteriophage P1 is an important tool for genetic engineering in mammalian cells. We constructed lentiviral vectors that efficiently deliver Cre in vitro and in vivo. Surprisingly, we found a significant reduction in proliferation and an accumulation in the G2/M phase of Cre-expressing cells. To minimize the toxic effect of Cre, we designed a lentiviral vector that integrates into the host genome, expresses Cre in the target cell, and is subsequently deleted from the genome in a Cre-dependent manner. Thus, the activity of Cre terminates its own expression (self-deleting). We showed efficient modification of target genes in vitro and in the brain after transduction with the self-deleting vectors. In contrast to sustained Cre expression, transient expression of Cre from the self-deleting vector induced significantly less cytotoxicity. Such a self-deleting Cre vector is a promising tool for the induction of conditional gene modifications with minimal Cre toxicity in vivo.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.201415498