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March 07, 2013

Japanese researchers have improved cloning for over 25 generations of cloning and normal lifespans

Using the technique that created Dolly the sheep, researchers from the RIKEN Center for Developmental Biology in Kobe, Japan have identified a way to produce healthy mouse clones that live a normal lifespan and can be sequentially cloned indefinitely.

In an experiment that started in 2005, the team led by Dr. Teruhiko Wakayama has used a technique called somatic cell nuclear transfer (SNCT) to produce 581 clones of one original 'donor' mouse, through 25 consecutive rounds of cloning.

By improving each step of the SCNT procedure, they were able to clone the mice repeatedly 25 times without seeing a reduction in the success rate. The 581 healthy mice obtained in this way were all fertile, they gave birth to healthy pups and lived a normal lifespan of about two years, similar to normally conceived mice.

“Our results show that there were no accumulations of epigenetic or genetic abnormalities in the mice, even after repeated cloning,” conclude the authors.

Dr. Wakayama adds, “This technique could be very useful for the large-scale production of superior-quality animals, for farming or conservation purposes.”

Successful Serial Recloning in the Mouse over Multiple Generations



Highlights

* Successful recloning of viable mice over 25 generations
* Cloning efficiency remains consistent over 25 iterations
* No evidence for the accumulation of reprogramming or genomic errors
* Serially recloned mice have the same characteristics as standard clones

Summary

Previous studies of serial cloning in animals showed a decrease in efficiency over repeated iterations and a failure in all species after a few generations. This limitation led to the suggestion that repeated recloning might be inherently impossible because of the accumulation of lethal genetic or epigenetic abnormalities. However, we have now succeeded in carrying out repeated recloning in the mouse through a somatic cell nuclear transfer method that includes a histone deacetylase inhibitor. The cloning efficiency did not decrease over 25 generations, and, to date, we have obtained more than 500 viable offspring from a single original donor mouse. The reprogramming efficiency also did not increase over repeated rounds of nuclear transfer, and we did not see the accumulation of reprogramming errors or clone-specific abnormalities. Therefore, our results show that repeated iterative recloning is possible and suggest that, with adequately efficient techniques, it may be possible to reclone animals indefinitely.


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