Difference between revisions of "Pye 2006 Nucleic Acids Research"
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{{Publication | {{Publication | ||
|title=Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN (2006) Production of transmitochondrial cybrids containing naturally occurring pathogenic mtDNA variants. Nucleic Acids Research 34: e95 | |title=Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN (2006) Production of transmitochondrial cybrids containing naturally occurring pathogenic mtDNA variants. Nucleic Acids Research 34:e95 | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/16885236 PMID: 16885236 Open Access] | |||
|authors=Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN | |authors=Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN | ||
|year=2006 | |year=2006 | ||
|journal=Nucleic Acids Res | |journal=Nucleic Acids Res | ||
|abstract=The human mitochondrial genome (mtDNA) | |abstract=The human mitochondrial genome (mtDNA) encodes polypeptides that are critical for coupling oxidative phosphorylation. Our detailed understanding of the molecular processes that mediate mitochondrial gene expression and the structure–function relationships of the OXPHOS components could be greatly improved if we were able to transfect mitochondria and manipulate mtDNA ''in vivo''. | ||
encodes polypeptides that are critical for coupling | Increasing our knowledge of this process is not merely of fundamental importance, as mutations of the mitochondrial genome are known to cause a | ||
oxidative phosphorylation. Our detailed understanding | spectrum of clinical disorders and have been implicated in more common neurodegenerative disease and the ageing process. In organellar or ''in vitro'' | ||
of the molecular processes that mediate | reconstitution studies have identified many factors central to the mechanisms of mitochondrial gene expression, but being able to investigate the molecular | ||
mitochondrial gene expression and the | aetiology of a limited number of cell lines from patients harbouring mutated mtDNA has been enormously beneficial. In the absence of a mechanism | ||
for manipulating mtDNA, a much larger pool of pathogenic mtDNA mutations would increase our knowledge of mitochondrial gene expression. | |||
could be greatly improved if we were able to transfect | Colonic crypts from ageing individuals harbour mutated mtDNA. Here we show that by generating cytoplasts from colonocytes, standard fusion techniques | ||
mitochondria and manipulate mtDNA in vivo. | can be used to transfer mtDNA into rapidly dividing immortalized cells and, thereby, respiratory-deficient transmitochondrial cybrids can be isolated. A simple screen identified clones that carried putative pathogenic mutations in MTRNR1, MTRNR2, MTCOI and MTND2, MTND4 and MTND6. | ||
Increasing our knowledge of this process is not | This method can therefore be exploited to produce a library of cell lines carrying pathogenic human mtDNA for further study. | ||
merely of fundamental importance, as mutations of | |mipnetlab=DE Munich Elstner M, UK Newcastle Lightowlers RN | ||
the mitochondrial genome are known to cause a | |discipline=Biomedicine | ||
spectrum of clinical disorders and have been implicated | |||
in more common neurodegenerative disease | |||
and the ageing process. In organellar or in vitro | |||
reconstitution studies have identified many factors | |||
central to the mechanisms of mitochondrial gene | |||
expression, but being able to investigate the molecular | |||
aetiology of a limited number of cell lines from | |||
patients harbouring mutated mtDNA has been | |||
enormously beneficial. In the absence of a mechanism | |||
for manipulating mtDNA, a much larger pool | |||
of pathogenic mtDNA mutations would increase | |||
our knowledge of mitochondrial gene expression. | |||
Colonic crypts from ageing individuals harbour | |||
mutated mtDNA. Here we show that by generating | |||
cytoplasts from colonocytes, standard fusion techniques | |||
can be used to transfer mtDNA into | |||
rapidly dividing immortalized cells and, thereby, | |||
respiratory-deficient transmitochondrial cybrids can | |||
be isolated. A simple screen identified clones that | |||
carried putative pathogenic mutations in MTRNR1, | |||
MTRNR2, MTCOI and MTND2, MTND4 and MTND6. | |||
This method can therefore be exploited to produce a | |||
library of cell lines carrying pathogenic human | |||
mtDNA for further study. | |||
| | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
| | |area=Respiration, mtDNA;mt-genetics, mt-Medicine | ||
| | |diseases=Cancer, Other | ||
|organism=Human | |organism=Human | ||
|preparations=Intact | |preparations=Intact cells | ||
| | |couplingstates=OXPHOS | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
| | |discipline=Biomedicine | ||
}} | }} | ||
Latest revision as of 13:42, 27 March 2018
| Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN (2006) Production of transmitochondrial cybrids containing naturally occurring pathogenic mtDNA variants. Nucleic Acids Research 34:e95 |
Pye D, Kyriakouli DS, Taylor GA, Johnson R, Elstner M, Meunier B, Chrzanowska-Lightowlers ZMA, Taylor RW, Turnbull DM, Lightowlers RN (2006) Nucleic Acids Res
Abstract: The human mitochondrial genome (mtDNA) encodes polypeptides that are critical for coupling oxidative phosphorylation. Our detailed understanding of the molecular processes that mediate mitochondrial gene expression and the structure–function relationships of the OXPHOS components could be greatly improved if we were able to transfect mitochondria and manipulate mtDNA in vivo. Increasing our knowledge of this process is not merely of fundamental importance, as mutations of the mitochondrial genome are known to cause a spectrum of clinical disorders and have been implicated in more common neurodegenerative disease and the ageing process. In organellar or in vitro reconstitution studies have identified many factors central to the mechanisms of mitochondrial gene expression, but being able to investigate the molecular aetiology of a limited number of cell lines from patients harbouring mutated mtDNA has been enormously beneficial. In the absence of a mechanism for manipulating mtDNA, a much larger pool of pathogenic mtDNA mutations would increase our knowledge of mitochondrial gene expression. Colonic crypts from ageing individuals harbour mutated mtDNA. Here we show that by generating cytoplasts from colonocytes, standard fusion techniques can be used to transfer mtDNA into rapidly dividing immortalized cells and, thereby, respiratory-deficient transmitochondrial cybrids can be isolated. A simple screen identified clones that carried putative pathogenic mutations in MTRNR1, MTRNR2, MTCOI and MTND2, MTND4 and MTND6. This method can therefore be exploited to produce a library of cell lines carrying pathogenic human mtDNA for further study.
• O2k-Network Lab: DE Munich Elstner M, UK Newcastle Lightowlers RN
Labels: MiParea: Respiration, mtDNA;mt-genetics, mt-Medicine
Pathology: Cancer, Other
Organism: Human
Preparation: Intact cells
Coupling state: OXPHOS
HRR: Oxygraph-2k
