DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor

Jill Glasspool-Malone, Peter R. Steenland, Ruth J. McDonald, Rigoberto A. Sanchez, Tammara Watts, Joseph Zabner, Robert W. Malone

Research output: Contribution to journalArticle

37 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)323-332
Number of pages10
JournalJournal of Gene Medicine
Volume4
Issue number3
DOIs
StatePublished - May 2002
Externally publishedYes

Fingerprint

Macaca
Transfection
Lung
Deoxyribonucleases
DNA
Primates
Microspheres
Technology Transfer
Hematoxylin
Eosine Yellowish-(YS)
Transgenes
Genes
Rodentia
Body Weight
Staining and Labeling

Keywords

  • ATA
  • Gene therapy
  • Lung
  • Naked DNA
  • Nuclease inhibitor
  • Transfection

ASJC Scopus subject areas

  • Genetics

Cite this

Glasspool-Malone, J., Steenland, P. R., McDonald, R. J., Sanchez, R. A., Watts, T., Zabner, J., & Malone, R. W. (2002). DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor. Journal of Gene Medicine, 4(3), 323-332. https://doi.org/10.1002/jgm.259

DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor. / Glasspool-Malone, Jill; Steenland, Peter R.; McDonald, Ruth J.; Sanchez, Rigoberto A.; Watts, Tammara; Zabner, Joseph; Malone, Robert W.

In: Journal of Gene Medicine, Vol. 4, No. 3, 05.2002, p. 323-332.

Research output: Contribution to journalArticle

Glasspool-Malone, J, Steenland, PR, McDonald, RJ, Sanchez, RA, Watts, T, Zabner, J & Malone, RW 2002, 'DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor', Journal of Gene Medicine, vol. 4, no. 3, pp. 323-332. https://doi.org/10.1002/jgm.259
Glasspool-Malone J, Steenland PR, McDonald RJ, Sanchez RA, Watts T, Zabner J et al. DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor. Journal of Gene Medicine. 2002 May;4(3):323-332. https://doi.org/10.1002/jgm.259
Glasspool-Malone, Jill ; Steenland, Peter R. ; McDonald, Ruth J. ; Sanchez, Rigoberto A. ; Watts, Tammara ; Zabner, Joseph ; Malone, Robert W. / DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor. In: Journal of Gene Medicine. 2002 ; Vol. 4, No. 3. pp. 323-332.
@article{253aa205ead74059b7935b607be5c902,
title = "DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor",
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.",
keywords = "ATA, Gene therapy, Lung, Naked DNA, Nuclease inhibitor, Transfection",
author = "Jill Glasspool-Malone and Steenland, {Peter R.} and McDonald, {Ruth J.} and Sanchez, {Rigoberto A.} and Tammara Watts and Joseph Zabner and Malone, {Robert W.}",
year = "2002",
month = "5",
doi = "10.1002/jgm.259",
language = "English (US)",
volume = "4",
pages = "323--332",
journal = "Journal of Gene Medicine",
issn = "1099-498X",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor

AU - Glasspool-Malone, Jill

AU - Steenland, Peter R.

AU - McDonald, Ruth J.

AU - Sanchez, Rigoberto A.

AU - Watts, Tammara

AU - Zabner, Joseph

AU - Malone, Robert W.

PY - 2002/5

Y1 - 2002/5

N2 - 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.

AB - 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.

KW - ATA

KW - Gene therapy

KW - Lung

KW - Naked DNA

KW - Nuclease inhibitor

KW - Transfection

UR - http://www.scopus.com/inward/record.url?scp=0036579881&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036579881&partnerID=8YFLogxK

U2 - 10.1002/jgm.259

DO - 10.1002/jgm.259

M3 - Article

VL - 4

SP - 323

EP - 332

JO - Journal of Gene Medicine

JF - Journal of Gene Medicine

SN - 1099-498X

IS - 3

ER -