NDLI: Role of endonuclease G in exogenous DNA stability in HeLa cells

Content Provider	Springer Nature Link
Author	Misic, V. El Mogy, M. Haj Ahmad, Y.
Copyright Year	2016
Abstract	Endonuclease G (EndoG) is a well-conserved mitochondrial-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four-day time course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances, it may nonspecifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from nonviral DNA vectors in gene therapy efforts.
Starting Page	163
Ending Page	175
Page Count	13
File Format	PDF
ISSN	00062979
Journal	Biochemistry (Moscow)
Volume Number	81
Issue Number	2
e-ISSN	16083040
Language	English
Publisher	Pleiades Publishing
Publisher Date	2016-02-16
Publisher Place	Moscow
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	EndoG HeLa cells plasmids shRNAs silencing transfection Biochemistry Bioorganic Chemistry Microbiology Biomedicine general
Content Type	Text
Resource Type	Article
Subject	Biochemistry Biophysics Geriatrics and Gerontology

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