DNA delivery to mitochondria: sequence specificity and energy enhancement
N Ibrahim, H Handa, A Cosset, M Koulintchenko… - Pharmaceutical …, 2011 - Springer
N Ibrahim, H Handa, A Cosset, M Koulintchenko, Y Konstantinov, RN Lightowlers, A Dietrich…
Pharmaceutical research, 2011•SpringerPurpose Mitochondria are competent for DNA uptake in vitro, a mechanism which may
support delivery of therapeutic DNA to complement organelle DNA mutations. We document
here key aspects of the DNA import process, so as to further lay the ground for mitochondrial
transfection in intact cells. Methods We developed DNA import assays with isolated
mitochondria from different organisms, using DNA substrates of various sequences and
sizes. Further import experiments investigated the possible role of ATP and protein …
support delivery of therapeutic DNA to complement organelle DNA mutations. We document
here key aspects of the DNA import process, so as to further lay the ground for mitochondrial
transfection in intact cells. Methods We developed DNA import assays with isolated
mitochondria from different organisms, using DNA substrates of various sequences and
sizes. Further import experiments investigated the possible role of ATP and protein …
Purpose
Mitochondria are competent for DNA uptake in vitro, a mechanism which may support delivery of therapeutic DNA to complement organelle DNA mutations. We document here key aspects of the DNA import process, so as to further lay the ground for mitochondrial transfection in intact cells.
Methods
We developed DNA import assays with isolated mitochondria from different organisms, using DNA substrates of various sequences and sizes. Further import experiments investigated the possible role of ATP and protein phosphorylation in the uptake process. The fate of adenine nucleotides and the formation of phosphorylated proteins were analyzed.
Results
We demonstrate that the efficiency of mitochondrial uptake depends on the sequence of the DNA to be translocated. The process becomes sequence-selective for large DNA substrates. Assays run with a natural mitochondrial plasmid identified sequence elements which promote organellar uptake. ATP enhances DNA import and allows tight integration of the exogenous DNA into mitochondrial nucleoids. ATP hydrolysis has to occur during the DNA uptake process and might trigger phosphorylation of co-factors.
Conclusions
Our data contribute critical information to optimize DNA delivery into mitochondria and open the prospect of targeting whole mitochondrial genomes or complex constructs into mammalian organelles in vitro and in vivo.
Springer
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