Journal article
DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor
The journal of gene medicine, Vol.4(3), pp.323-332
05/2002
DOI: 10.1002/jgm.259
PMID: 12112649
Abstract
Background
Nuclease activity present within respiratory tissues contributes to the rapid clearance of injected DNA and therefore may reduce the transfection activity of directly injected transgenes. Most gene transfer technologies transduce or transfect murine tissues more efficiently than corresponding primate tissues. Therefore, it is prudent to assess the utility of novel gene transfer strategies in both rodent and primate models before proceeding with further development.
Methods
This study analyzed the effects of ATA (a nuclease inhibitor) on the direct transfection of macaque and murine lung tissue; compared the levels of DNase activity in murine, primate, and human lung fluids; and tested the inhibitory activity of ATA on the DNase activity present in these samples. Fluorescent microspheres were used to detect areas of transfection in lung.
Results
Intratracheal administration of a nuclease inhibitor (ATA) with naked DNA (0.5 µg ATA/g body weight) enhanced direct transfection efficacy in macaque lung by over 86-fold and by over 54-fold in mouse lung. Hematoxylin and eosin staining showed no apparent tissue toxicity. Moreover, macaque, human, and mouse lung fluids were found to possess similar levels of DNase activity and this activity was inhibited by similar concentrations of ATA. The authors also successfully pioneered the use of carboxylate-modified microsphere tracers to identify areas of transfection and/or treatment.
Conclusion
This work provides evidence that using direct nuclease inhibitors will enhance lung transfection and that nuclease activity is present in all lung fluids tested, which can be inhibited by the use of direct DNase inhibitors.
Details
- Title: Subtitle
- DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor
- Creators
- Jill Glasspool-Malone - Gene Delivery Alliance, Inc., Rockville, MD 20850, USAPeter R Steenland - Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USARuth J McDonald - California Regional Primate Research Center, University of California at Davis, Davis, CA, USARigoberto A Sanchez - Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USATammara L Watts - Center for Vaccine Development at the University of Maryland School of Medicine, Baltimore, MD, USAJoseph Zabner - University of Iowa, IA, USARobert W Malone - Gene Delivery Alliance, Inc., Rockville, MD 20850, USA
- Resource Type
- Journal article
- Publication Details
- The journal of gene medicine, Vol.4(3), pp.323-332
- Publisher
- John Wiley & Sons, Ltd
- DOI
- 10.1002/jgm.259
- PMID
- 12112649
- ISSN
- 1099-498X
- eISSN
- 1521-2254
- Number of pages
- 10
- Language
- English
- Date published
- 05/2002
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
- Record Identifier
- 9984094378502771
Metrics
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