Methods for Efficient Elimination of Mitochondrial DNA from Cultured Cells

被引：27

作者：

Spadafora, Domenico ^{[1
]}

Kozhukhar, Nataliya ^{[2
]}

Chouljenko, Vladimir N. ^{[3
,4
]}

Kousoulas, Konstantin G. ^{[3
,4
]}

Alexeyev, Mikhail F. ^{[2
,5
]}

机构：

[1] Univ S Alabama, Dept Pharmacol, Mobile, AL 36688 USA

[2] Univ S Alabama, Dept Physiol & Cell Biol, Mobile, AL 36688 USA

[3] Louisiana State Univ, Sch Vet Med, Div Biotechnol & Mol Med, Baton Rouge, LA 70803 USA

[4] Louisiana State Univ, Sch Vet Med, Dept Pathobiol Sci, Baton Rouge, LA 70803 USA

[5] Univ S Alabama, Ctr Lung Biol, Mobile, AL 36688 USA

来源：

PLOS ONE | 2016年 / 11卷 / 05期

基金：

美国国家卫生研究院;

关键词：

HEREDITARY OPTIC NEUROPATHY; XENOMITOCHONDRIAL CYBRIDS; GENE-EXPRESSION; SEQUENCE; MTDNA; ORGANIZATION; DEGRADATION; INHERITANCE; BIOGENESIS; TERMINUS;

D O I：

10.1371/journal.pone.0154684

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Here, we document that persistent mitochondria DNA (mtDNA) damage due to mitochondrial overexpression of the Y147A mutant uracil-N-glycosylase as well as mitochondrial overexpression of bacterial Exonuclease III or Herpes Simplex Virus protein UL12.5M185 can induce a complete loss of mtDNA (rho(0) phenotype) without compromising the viability of cells cultured in media supplemented with uridine and pyruvate. Furthermore, we use these observations to develop rapid, sequence-independent methods for the elimination of mtDNA, and demonstrate utility of these methods for generating rho(0) cells of human, mouse and rat origin. We also demonstrate that rho(0) cells generated by each of these three methods can serve as recipients of mtDNA in fusions with enucleated cells.

引用

页数：18

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