A vector-host system to fingerprint virus tropism.

Matthew L. Hillestad, Adam J. Guenzel, Karl A Nath, Michael A Barry

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Reporter genes are important tools for assessing vector pharmacology in vivo. Although useful, current systems are limited by (1) the need to generate a new vector for each different reporter, (2) the inability to package reporter genes in small vectors, and (3) variations in reporter gene feedback due to variations in cell-to-cell vector copy number. To circumvent these problems, we have used Cre recombinase as a "cat's paw" to activate reporter genes embedded in transgenic mice. The small Cre gene was introduced into self-complementary adeno-associated viral (scAAV) vectors with limited packaging capacity. Injection of scAAV-Cre vectors into mice with loxP-inactivated luciferase enabled in vivo imaging distributions comparable to the signal observed after AAV-luciferase injection. When injected into mT/mG mice, AAV-Cre converted ubiquitous expression of red fluorescent protein (RFP) to green fluorescent protein (GFP) expression only where the vectors transduced cells. Injection into F(1) hybrid luciferase and mT/mG mice enabled simultaneous three-reporter tracking. This system was able to discriminate cell-specific transduction in all organs tested, with particular usefulness for detecting AAV serotype-specific transduction in the liver, kidney, and muscle. Given that F(1) mice bear exactly one copy of luciferase and one copy of RFP-GFP, each reporter gene is either "on" or "off" in a cell. The Cre system therefore provides a unique quantum method to quantify vector delivery that can be applied when vector capacity is limited.

Original languageEnglish (US)
Pages (from-to)1116-1126
Number of pages11
JournalHuman Gene Therapy
Volume23
Issue number10
StatePublished - Oct 2012

Fingerprint

Tropism
Dermatoglyphics
Reporter Genes
Luciferases
Viruses
Green Fluorescent Proteins
Injections
Product Packaging
Transgenic Mice
Cats
Pharmacology
Kidney
Muscles
Liver
Genes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

A vector-host system to fingerprint virus tropism. / Hillestad, Matthew L.; Guenzel, Adam J.; Nath, Karl A; Barry, Michael A.

In: Human Gene Therapy, Vol. 23, No. 10, 10.2012, p. 1116-1126.

Research output: Contribution to journalArticle

Hillestad, ML, Guenzel, AJ, Nath, KA & Barry, MA 2012, 'A vector-host system to fingerprint virus tropism.', Human Gene Therapy, vol. 23, no. 10, pp. 1116-1126.
Hillestad, Matthew L. ; Guenzel, Adam J. ; Nath, Karl A ; Barry, Michael A. / A vector-host system to fingerprint virus tropism. In: Human Gene Therapy. 2012 ; Vol. 23, No. 10. pp. 1116-1126.
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