This Article Full Text Full Text (PDF) Supplemental material Other Versions of this Article: JVI.00623-08v1 82/14/7111    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Google Scholar Articles by Sauvain, M.-O. Articles by Trono, D. PubMed PubMed Citation Articles by Sauvain, M.-O. Articles by Trono, D.

 Previous Article  |  Next Article 

Journal of Virology, July 2008, p. 7111-7119, Vol. 82, No. 14
0022-538X/08/$08.00+0     doi:10.1128/JVI.00623-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genotypic Features of Lentivirus Transgenic Mice ^,

Marc-Olivier Sauvain,¹ Alexander P. Dorr,^1, Brian Stevenson,² Alexandra Quazzola,¹ Félix Naef,¹ Maciej Wiznerowicz,¹ Frédéric Schütz,² Victor Jongeneel,² Denis Duboule,^1,3 François Spitz,^3, and Didier Trono^1*

School of Life Sciences and Frontiers in Genetics National Center for Competence in Research, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland,¹ Ludwig Institute for Cancer Research and Swiss Institute of Bioinformatics, 1015 Lausanne, Switzerland,² Department of Zoology and Animal Biology, University of Geneva, Geneva, Switzerland³

Received 20 March 2008/ Accepted 30 April 2008

Lentivector-mediated transgenesis is increasingly used, whether for basic studies as an alternative to pronuclear injection of naked DNA or to test candidate gene therapy vectors. In an effort to characterize the genetic features of this approach, we first measured the frequency of germ line transmission of individual proviruses established by infection of fertilized mouse oocytes. Seventy integrants from 11 founder (G0) mice were passed to 111 first generation (G1) pups, for a total of 255 events corresponding to an average rate of transmission of 44%. This implies that integration had most often occurred at the one- or two-cell stage and that the degree of genotypic mosaicism in G0 mice obtained through this approach is generally minimal. Transmission analysis of eight individual proviruses in 13 G2 mice obtained by a G0-G1 cross revealed only 8% of proviral homozygosity, significantly below the 25% expected from purely Mendelian transmission, suggesting counter-selection due to interference with the functions of targeted loci. Mapping of 239 proviral integration sites in 49 founder animals revealed that about 60% resided within annotated genes, with a marked tendency for clustering in the middle of the transcribed region, and that integration was not influenced by the transcriptional orientation. Transcript levels of a set of arbitrarily chosen target genes were significantly higher in two-cell embryos than in embryonic stem cells or adult somatic cells, suggesting that, as previously noted in other settings, lentiviral vectors integrate preferentially into regions of the genome that are transcriptionally active or poised for activation.

Published ahead of print on 7 May 2008.

Supplemental material for this article may be found at http://jvi.asm.org/.

Present address: Merck Serono International S.A., 9 Chemin des Mines, 1202 Geneva, Switzerland.

Present address: EMBL, Meyerhofstr. 1, 69117 Heidelberg, Germany.