HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 293/1/H881    most recent 00060.2007v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (6) Citing Articles via Google Scholar Google Scholar Articles by Michalkiewicz, M. Articles by Cowley, A. W. Search for Related Content PubMed PubMed Citation Articles by Michalkiewicz, M. Articles by Cowley, A. W., Jr.

INNOVATIVE METHODOLOGY

Efficient transgenic rat production by a lentiviral vector

¹Department of Physiology, ²Human Molecular Genetic Center, and ⁴Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, Wisconsin; and ³Laboratory of Genetics, The Salk Institute, La Jolla, California

Submitted 15 January 2007 ; accepted in final form 20 February 2007

A lentiviral construct for an enhanced green fluorescent protein (eGFP) driven by a chicken -actin promoter, cytomegalovirus enhancer, and intronic sequences from rabbit -globin (CAG) was used to produce transgenic lines of rats for evaluation of the usefulness of this approach in gene function studies. Fertilized eggs were collected from inbred Dahl S and outbred Sprague-Dawley rats, and 100 pl of concentrated virus were microinjected into the perivitrelline space of one-cell embryos. Of 121 embryos injected, 60 pups (49.6%) were born. Transgenic rates averaged 22% in Dahl S and 14% in Sprague-Dawley rats. Copy number ranged from one to four in the founders, and the inheritance of the transgene in a subsequent F₁ population was 48.2%. The small number of insertion sites enabled us to derive inbred transgenic lines with a single copy of the transgene within one generation. Sequencing of each transgene insertion site revealed that they inserted as single copies with a preference for the introns of genes. The CAG promoter drove high levels of eGFP expression in brain, kidney, heart, and vasculature, making it very suitable for exploring the cardiovascular function of newly discovered genes. The pattern of eGFP expression was similar across five different F₁ transgenic lines, indicating that the expression of the transgene was independent of its chromosomal position. Thus lentiviral transgenesis provides a powerful tool for the production of transgenic inbred rats and will enhance the usefulness of this species in gene discovery and target validation studies.

gene targeting; transgenesis; lentiviral gene transfer; gene expression

This article has been cited by other articles:

				A. M. Geurts, G. J. Cost, Y. Freyvert, B. Zeitler, J. C. Miller, V. M. Choi, S. S. Jenkins, A. Wood, X. Cui, X. Meng, et al. Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases Science, July 24, 2009; 325(5939): 433 - 433. [Abstract] [Full Text] [PDF]

				F. Park Lentiviral vectors: are they the future of animal transgenesis? Physiol Genomics, October 19, 2007; 31(2): 159 - 173. [Abstract] [Full Text] [PDF]