Uptake of foreign nucleic acids in kidney tubular epithelial cells deficient in proapoptotic endonucleases

Timea Buzder, Xiaoyan Yin, Xiaoying Wang, Gaspar Banfalvi, Alexei G. Basnakian

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Degradation of DNA during gene delivery is an obstacle for gene transfer and for gene therapy. DNases play a major role in degrading foreign DNA. However, which of the DNases are involved and whether their inactivation can improve gene delivery have not been studied. We have recently identified deoxyribonuclease I (DNase I) and endonuclease G (EndoG) as the major degradative enzymes in the mouse kidney proximal tubule epithelial (TKPTS) cells. In this study, we used immortalized mouse TKPTS cells and primary tubular epithelial cells isolated from DNase I or EndoG knockout (KO) mice and examined the degradation of plasmid DNA during its uptake. DNase I and EndoG KO cells showed a higher rate of transfection by pECFP-N1 plasmid than wild-type cells. In addition, EndoG KO cells prevented the uptake of fluorescent-labeled RNA. Complete inhibition of secreted DNase I by G-actin did not improve plasmid transfection, indicating that only intracellular DNase I affects DNA stability. Data demonstrate the importance of DNase I and EndoG in host cell defense against gene and RNA delivery to renal tubular epithelial cells in vitro.

Original languageEnglish
Pages (from-to)435-442
Number of pages8
JournalDNA and Cell Biology
Volume28
Issue number9
DOIs
Publication statusPublished - Sep 1 2009

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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