Toward genetic transformation of mitochondria in mammalian cells using a recoded drug-resistant selection marker

Young Geol Yoon; Michael Duane Koob

doi:10.1016/j.jgg.2011.03.005

Volume 38 Issue 4

Apr. 2011

Turn off MathJax

Article Contents

Abstract

References

Article Navigation > Journal of Genetics and Genomics > 2011 > 38(4): 173-179

PDF( 419 KB)

Toward genetic transformation of mitochondria in mammalian cells using a recoded drug-resistant selection marker

doi: 10.1016/j.jgg.2011.03.005

Young Geol Yoon^a,
Michael Duane Koob^b

a
Mitochondria Hub Regulation Center and Department of Anatomy and Cell Biology, Dong-A University College of Medicine, 3-1 Dongdaesin-dong, Seo-gu, Busan 602-714, Republic of Korea
b
Department of Laboratory Medicine and Pathology and Institute of Human Genetics, University of Minnesota Medical School, 420 Delaware St. S.E., Minneapolis, MN 55455, USA

More Information

Corresponding author: E-mail address: youngyoon97@gmail.com (Young Geol Yoon)
Received Date: 2010-12-10
Accepted Date: 2011-03-10
Rev Recd Date: 2011-03-07

Available Online: 2011-03-23

Publish Date: 2011-04-20

Abstract

Abstract

Due to technical difficulties, the genetic transformation of mitochondria in mammalian cells is still a challenge. In this report, we described our attempts to transform mammalian mitochondria with an engineered mitochondrial genome based on selection using a drug resistance gene. Because the standard drug-resistant neomycin phosphotransferase confers resistance to high concentrations of G418 when targeted to the mitochondria, we generated a recoded neomycin resistance gene that uses the mammalian mitochondrial genetic code to direct the synthesis of this protein in the mitochondria, but not in the nucleus (mitochondrial version). We also generated a universal version of the recoded neomycin resistance gene that allows synthesis of the drug-resistant proteins both in the mitochondria and nucleus. When we transfected these recoded neomycin resistance genes that were incorporated into the mouse mitochondrial genome clones into mouse tissue culture cells by electroporation, no DNA constructs were delivered into the mitochondria. We found that the universal version of the recoded neomycin resistance gene was expressed in the nucleus and thus conferred drug resistance to G418 selection, while the synthetic mitochondrial version of the gene produced no background drug-resistant cells from nuclear transformation. These recoded synthetic drug-resistant genes could be a useful tool for selecting mitochondrial genetic transformants as a precise technology for mitochondrial transformation is developed.
- Mitochondrial codon,
- Recode,
- Mitochondrial transformation,
- Selection marker,
- G418

FullText(HTML)

References(28)

References

[1]	Andersson, G.E., Kurland, C.G. An extreme codon preference strategy: codon reassignment Mol. Biol. Evol., 8 (1991),pp. 530-544
[2]	Barrientos, A., Barros, M.H., Valnot, I. et al. Cytochrome oxidase in health and disease Gene, 286 (2002),pp. 53-63
[3]	Barry, P., Pratt-Lowe, E., Luciw, P.A. Electroporation of T-cell and macrophage cell lines Bio-Rad Lab. Tech. Bull., 1349 (1989)
[4]	Boynton, J.E., Gillham, N.W. Methods Enzymol., 264 (1996),pp. 279-296
[5]	Butow, R.A., Henke, R.M., Moran, J.V. et al. Methods Enzymol., 264 (1996),pp. 265-278
[6]	Delorme, M.O., Henaut, A. Codon usage is imposed by the gene location in the transcription unit Curr. Genet., 20 (1991),pp. 353-358
[7]	DiMauro, S., Schon, E.A. Mitochondrial DNA mutations in human disease Am. J. Med. Genet., 106 (2001),pp. 18-26
[8]	Fox, T.D., Sanford, J.C., McMullin, T.W. Plasmids can stably transform yeast mitochondria lacking endogenous mtDNA Proc. Natl. Acad. Sci. USA, 85 (1988),pp. 7288-7292
[9]	Gilkerson, R.W., Schon, E.A., Hernandez, E. et al. Mitochondrial nucleoids maintain genetic autonomy but allow for functional complementation J. Cell Biol., 181 (2008),pp. 1117-1128
[10]	Irwin, M.H., Johnson, L.W., Pinkert, C.A. Isolation and microinjection of somatic cell-derived mitochondria and germline heteroplasmy in transmitochondrial mice Transgenic Res., 8 (1999),pp. 119-123
[11]	Johnston, S.A., Anziano, P.Q., Shark, K. et al. Mitochondrial transformation in yeast by bombardment with microprojectiles Science, 240 (1988),pp. 1538-1541
[12]	Kmiec, B., Woloszynska, M., Janska, H. Heteroplasmy as a common state of mitochondrial genetic information in plants and animals Curr. Genet., 50 (2006),pp. 149-159
[13]	Montoya, J., Ojala, D., Attardi, G. Distinctive features of the 5′-terminal sequences of the human mitochondrial mRNAs Nature, 290 (1981),pp. 465-470
[14]	Pecina, P., Houstkova, H., Hansikova, H. et al. Genetic defects of cytochrome c oxidase assembly Physiol. Res., 53 (2004),pp. S213-S223
[15]	Potter, H. Gene transfer by electroporation Bio-Rad Lab. Tech. Bull., 1346 (1988)
[16]	Rustenbeck, I., Eggers, G., Reiter, H. et al. Polyamine modulation of mitochondrial calcium transport. I. Stimulatory and inhibitory effects of aliphatic polyamines, aminoglucosides and other polyamine analogues on mitochondrial calcium uptake Biochem. Pharmacol., 56 (1998),pp. 977-985
[17]	Saraste, M. Science, 283 (1999),pp. 1488-1493
[18]	Schon, E.A., Gilkerson, R.W. Functional complementation of mitochondrial DNAs: mobilizing mitochondrial genetics against dysfunction Biochim. Biophys. Acta, 1800 (2010),pp. 245-249
[19]	Takeda, K., Tasai, M., Iwamoto, M. et al. Microinjection of cytoplasm or mitochondria derived from somatic cells affects parthenogenetic development of murine oocytes Biol. Reprod., 72 (2005),pp. 1397-1404
[20]	Trifunovic, A., Wredenberg, A., Falkenberg, M. et al. Premature ageing in mice expressing defective mitochondrial DNA polymerase Nature, 429 (2004),pp. 417-423
[21]	Wallace, D.C. Mitochondrial diseases in man and mouse Science, 283 (1999),pp. 1482-1488
[22]	Wallace, D.C. A mitochondrial paradigm for degenerative diseases and ageing Novartis Found. Symp., 235 (2001),pp. 247-263
[23]	Williams, B.A., Hirt, R.P., Lucocq, J.M. et al. Nature, 418 (2002),pp. 865-869
[24]	Yoon, Y.G., Koob, M.D. Nucleic Acids Res., 31 (2003),pp. 1407-1415
[25]	Yoon, Y.G., Koob, M.D. Transformation of isolated mammalian mitochondria by bacterial conjugation Nucleic Acids Res., 33 (2005),p. e139
[26]	Yoon, Y.G., Koob, M.D. Selection by drug resistance proteins located in the mitochondria of mammalian cells Mitochondrion, 8 (2008),pp. 345-351
[27]	Yoon, Y.G., Koob, M.D., Yoo, Y.H. Re-engineering the mitochondrial genomes in mammalian cells Anat. Cell Biol., 43 (2010),pp. 97-109
[28]	Yoon, Y.G., Yang, Y.W., Koob, M.D. Biotechnol. Lett., 31 (2009),pp. 1671-1676

Relative Articles

[1]	Chen Qu, Yating Kan, Hui Zuo, Mengqi Wu, Zhixiang Dong, Xinyi Wang, Qing Zhang, Heng Wang, Dou Wang, Jiong Chen. Actin polymerization induces mitochondrial distribution during collective cell migration[J]. Journal of Genetics and Genomics, 2023, 50(1): 46-49. doi: 10.1016/j.jgg.2022.04.014
[2]	Zhejun Ji, Guang-Hui Liu, Jing Qu. Mitochondrial sirtuins, metabolism, and aging[J]. Journal of Genetics and Genomics, 2022, 49(4): 287-298. doi: 10.1016/j.jgg.2021.11.005
[3]	Yu Sun, Xiujuan Wei, Fang Fang, Yiping Shen, Haiyan Wei, Jiuwei Li, Xianglai Ye, Yongkun Zhan, Xiantao Ye, Xiaomin Liu, Wei Yang, Yuhua Li, Xiangju Geng, Xuelin Huang, Yiyan Ruan, Zailong Qin, Shang Yi, Jianxin Lyu, Hezhi Fang, Yongguo Yu. HPDL deficiency causes a neuromuscular disease by impairing the mitochondrial respiration[J]. Journal of Genetics and Genomics, 2021, 48(8): 727-736. doi: 10.1016/j.jgg.2021.01.009
[4]	Hieu Nguyen, Thomas LaFramboise. Complexities and pitfalls in analyzing and interpreting mitochondrial DNA content in human cancer[J]. Journal of Genetics and Genomics, 2020, 47(7): 349-359. doi: 10.1016/j.jgg.2020.04.007
[5]	Ammar Husami, Jesse Slone, Jenice Brown, Meghan Bromwell, C. Alexander Valencia, Taosheng Huang. Clinical utility of whole genome sequencing for the detection of mitochondrial genome mutations[J]. Journal of Genetics and Genomics, 2020, 47(3): 167-169. doi: 10.1016/j.jgg.2020.03.001
[6]	Ruofeng Tang, Xin Wang, Junxiang Zhou, Fengxia Zhang, Shan Zhao, Qiwen Gan, Liyuan Zhao, Fengyang Wang, Qian Zhang, Jie Zhang, Guodong Wang, Chonglin Yang. Defective arginine metabolism impairs mitochondrial homeostasis in Caenorhabditis elegans[J]. Journal of Genetics and Genomics, 2020, 47(3): 145-156. doi: 10.1016/j.jgg.2020.02.007
[7]	Zhe Chen, Fan Zhang, Hong Xu. Human mitochondrial DNA diseases and Drosophila models[J]. Journal of Genetics and Genomics, 2019, 46(4): 201-212. doi: 10.1016/j.jgg.2019.03.009
[8]	Jesse Slone, Baoheng Gui, Taosheng Huang. The current landscape for the treatment of mitochondrial disorders[J]. Journal of Genetics and Genomics, 2018, 45(2): 71-77. doi: 10.1016/j.jgg.2017.11.008
[9]	Adeniyi C. Adeola, Sheila C. Ommeh, Jiao-Jiao Song, S. Charles Olaogun, Oscar J. Sanke, Ting-Ting Yin, Guo-Dong Wang, Shi-Fang Wu, Zhong-Yin Zhou, Jacqueline K. Lichoti, Bernard R. Agwanda, Philip M. Dawuda, Robert W. Murphy, Min-Sheng Peng, Ya-Ping Zhang. A cryptic mitochondrial DNA link between North European and West African dogs[J]. Journal of Genetics and Genomics, 2017, 44(3): 163-170. doi: 10.1016/j.jgg.2016.10.008
[10]	Xiang-Hong Ou, Qing-Yuan Sun. Mitochondrial replacement techniques or therapies (MRTs) to improve embryo development and to prevent mitochondrial disease transmission[J]. Journal of Genetics and Genomics, 2017, 44(8): 371-374. doi: 10.1016/j.jgg.2017.07.003
[11]	Basdeo Bhagwat, Ming Chi, Li Su, Haifeng Tang, Guiliang Tang, Yu Xiang. An in vivo Transient Expression System Can Be Applied for Rapid and Effective Selection of Artificial MicroRNA Constructs for Plant Stable Genetic Transformation[J]. Journal of Genetics and Genomics, 2013, 40(5): 261-270. doi: 10.1016/j.jgg.2013.03.012
[12]	Shengxin Chang, Jianmei Chen, Yankun Wang, Bingchao Gu, Jianbo He, Pu Chu, Rongzhan Guan. The Mitochondrial Genome of Raphanus sativus and Gene Evolution of Cruciferous Mitochondrial Types[J]. Journal of Genetics and Genomics, 2013, 40(3): 117-126. doi: 10.1016/j.jgg.2013.01.003
[13]	Laura L. Clay Montier, Janice J. Deng, Yidong Bai. Number matters: control of mammalian mitochondrial DNA copy number[J]. Journal of Genetics and Genomics, 2009, 36(3): 125-131. doi: 10.1016/S1673-8527(08)60099-5
[14]	Yu Ding, Yongyan Li, Junyan You, Li Yang, Bobei Chen, Jianxin Lu, Min-Xin Guan. Mitochondrial tRNAGlu A14693G variant may modulate the phenotypic manifestation of deafness-associated 12S rRNA A1555G mutation in a Han Chinese family[J]. Journal of Genetics and Genomics, 2009, 36(4): 241-250. doi: 10.1016/S1673-8527(08)60111-3
[15]	Qifa Li, Zhenshan Liu, Yinxia Li, Xingbo Zhao, Liyan Dong, Zengxiang Pan, Yuanrong Sun, Ning Li, Yinxue Xu, Zhuang Xie. Origin and phylogenetic analysis of Tibetan Mastiff based on the mitochondrial DNA sequence[J]. Journal of Genetics and Genomics, 2008, 35(6): 335-340. doi: 10.1016/S1673-8527(08)60049-1
[16]	Jinghua Yang, Mingfang Zhang, Jingquan Yu. Mitochondrial retrograde regulation tuning fork in nuclear genes expressions of higher plants[J]. Journal of Genetics and Genomics, 2008, 35(2): 65-71. doi: 10.1016/S1673-8527(08)60010-7
[17]	Hongying Qu, Fei Ma, Qingwei Li. Comparative analysis of mitochondrial fragments transferred to the nucleus in vertebrate[J]. Journal of Genetics and Genomics, 2008, 35(8): 485-490. doi: 10.1016/S1673-8527(08)60066-1
[18]	Wanshi Cai, Qun Fu, Xiangtian Zhou, Jia Qu, Yi Tong, Min-Xin Guan. Mitochondrial variants may influence the phenotypic manifestation of Leber's hereditary optic neuropathy-associated ND4 G11778A mutation[J]. Journal of Genetics and Genomics, 2008, 35(11): 649-655. doi: 10.1016/S1673-8527(08)60086-7
[19]	Shuqing Xu, Jiangbai Luosang, Sang Hua, Jian He, Asan Ciren, Wei Wang, Xiaomei Tong, Yu Liang, Jian Wang, Xiaoguang Zheng. High Altitude Adaptation and Phylogenetic Analysis of Tibetan Horse Based on the Mitochondrial Genome[J]. Journal of Genetics and Genomics, 2007, 34(8): 720-729. doi: 10.1016/S1673-8527(07)60081-2
[20]	Min Zhang, Tianwen Cao, Rui Zhang, Yaping Guo, Yihao Duan, Enbo Ma. Phylogeny of Apaturinae Butterflies (Lepidoptera: Nymphalidae) Based on Mitochondrial Cytochrome Oxidase? Gene[J]. Journal of Genetics and Genomics, 2007, 34(9): 812-823. doi: 10.1016/S1673-8527(07)60092-7