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 G(2)/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.
Pfeifer
A
A
The Salk Institute, La Jolla, CA 92037, USA.
Brandon
E P
EP
Kootstra
N
N
Gage
F H
FH
Verma
I M
IM
D000818:Animals; D001854:Bone Marrow Cells / Q000201:enzymology; D001921:Brain / Q000201:enzymology; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D005822:Genetic Vectors; D019426:Integrases / Q000235:genetics*; D016086:Lentivirus / Q000201:enzymology*; D051379:Mice; D008815:Mice, Inbred Strains; D014764:Viral Proteins / Q000235:genetics*
D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases