Mitochondria DNA and the organellar ribosomes differ from the chromosomal DNA and cytosolic ribosomes in several aspects, reflecting their prokaryotic origin in evolution. The similarity between mitochondrial biogenesis and bacterial/viral replication represents a significant challenge for drug developers designing novel antibacterials and antivirals, as many such drugs can cause serious mitochondrial toxicity.
This problem is now sufficiently well-recognized that the FDA in 2006 issued a guidance memo stating that new antiviral drugs should be screened for their potential mitochondrial toxicity. The Agency's position was reinforced in 2009 with the release of a guidance memo on Drug-Induced Liver Injury, in which the antiviral FIAU was cited as an example of the potential for antivirals to cause organ damage as a result of their toxic effects on mitochondria.
Figure 1. The graph illustrates rapid screening to identify antibiotics that inhibit mitochondrial biogenesis in vitro. Human HepG2 cells were grown in the indicated antibiotics for 5 population doublings and then effects on biogenesis were measured. Antibiotics that interfere with protein synthesis on mitochondrial ribosomes (Linezolid, Chloramphenicol, Tetracycline, Doxycycline and Minocycline) strongly inhibit mitochondrial biogenesis while other antibiotics do no not.
Measuring RNA levels using reverse transcription qPCR is occasionally discussed as a useful high-throughput technique for identifying effects on mitochondrial biogenesis [3], but this approach has a fundamental limitation, that being its inability to identify inhibition of mitochondrial ribosomes. If such an effect is occuring then RNA levels might in fact increase in the cell, since mitochondrial translation is unable to process transcription products. The only way to easily and simultaneously identify inhibition of both transcription and translation is to ratio the levels of an mtDNA-encoded protein with total cell protein and/or a nDNA-encoded protein. This can be done by Western blotting or by ICC microscopy, but neither technique is suitably quantitative, nor are they amenable to high-throughput nor clinical applications.
MitoSciences offers the only solution for measuring drug-induced effects on mitochondrial biogenesis early in the safety screening process. Our MitoBiogenesis™ In-Cell ELISA is a true duplexing 96/384-well assay that ratios both an mtDNA- and an nDNA-encoded protein in cultured or primary cells, and which requires very little sample prep and few overall steps.
Mitochondrial Biogenesis Assays
| Cat. No. | Name | Reactivity | Amount | Price | |
| MS642 | MitoBiogenesis™ In-Cell ELISA Kit (IR) | human, rat, mouse, bovine | 2 x 96 tests | $425.00 | |
| MS643 | MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) | human, rat, mouse, bovine | 2 x 96 tests | $425.00 |
