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New technology for an old favorite: lentiviral transgenesis and RNAi in rats

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Abstract

The ability to produce targeted deletions in the mouse genome via homologous recombination has been a hallmark of mouse genetics, and has lead to the production of thousands of gene knockouts. New technologies are making it possible to disrupt gene function in many other species. This article reviews some of these methods, highlighting the powerful combination of lentiviral vectors with RNA interference (RNAi), which allows one to produce transgenic animals expressing short hairpin RNA (shRNA) to “knock down” specific gene expression. Lentiviral transduction of embryos has been shown to be a highly efficient means of transgenesis, and is particularly promising for animals that are considered difficult to genetically modify by DNA pronuclear injection. This technique has been popular for introducing transgenes for shRNA expression into rodents and its utility for creating new genetic models has already been demonstrated. One of the purported advantages of in vivo RNAi is that shRNA expressing transgenes would be expected to act in a dominant nature, resulting in a phenotype in founder animals. However, one possible concern with lentiviral-mediated transgenesis is the potential for mosaicism in founders, and the data for this phenomenon and the potential causes and solutions are discussed. Emphasis is placed on the application of in vivo RNAi, and other reverse genetic methods, for creating new genetic models in the rat.

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Acknowledgements

This manuscript is written in memory of David Garbers, who was a caring mentor with an inspiring vision for the development of new technologies and a dream for discovery. He is deeply missed. Thank you also to Alma Alvarado for assistance in obtaining the unpublished data presented here, and Robert Hammer and Matthew Porteus for guidance and reading the manuscript. This work was supported by the Lalor Foundation, Inc., the Howard Hughes Medical Institute, and the Cecil H. and Ida Green Center for Reproductive Biology Sciences.

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  1. Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Room ND11.136, Dallas, TX, 75390-9051, USA

    Christina Tenenhaus Dann

  2. Cecil H. & Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Room ND11.136, Dallas, TX, 75390-9051, USA

    Christina Tenenhaus Dann

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Correspondence to Christina Tenenhaus Dann.

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Dann, C.T. New technology for an old favorite: lentiviral transgenesis and RNAi in rats. Transgenic Res 16, 571–580 (2007). https://doi.org/10.1007/s11248-007-9121-z

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