Researchers get viable mice by editing DNA from two sperm

Rare success

The researchers spent time ensuring that the enzymes they had were modifying the methylation as expected, and that development started as usual. Their general finding is that the enzymes did change the methylation state for about 500 bases on either side of the targeted site, and did so pretty consistently. But there are seven different imprinting sites that need to be modified, each of which controls multiple nearby genes. So, while the modifications were consistent, they weren’t always thorough enough to result in the expected changes to all of the nearby genes.

This limited efficiency showed up in the rate of survival. Starting with over 250 reprogrammed embryos that carried DNA from two males, they ended up with sixteen pregnancies, but only four that died at birth, and three live ones; based on other experiments, most of the rest died during the second half of embryonic development. Of the three live ones, one was nearly 40 percent larger than the typical pup, suggesting problems regulating growth—it died the day after birth.

All three live births were male, although the numbers are small enough that it’s impossible to tell if that’s significant or not.

The researchers suggest several potential reasons for the low efficiency. One is simply that, while the probability of properly reprogramming at least one of the sites is high, reprogramming all seven is considerably more challenging. There’s also the risk of off-target effects, where the modification takes place in locations with similar sequences to the ones targeted. They also concede that there could be other key imprinted regions that we simply haven’t identified yet.

Sorting that out would be needed if we want to use this approach as a tool, which might be potentially useful as a way to breed mice that carry mutations that affect female viability or fertility. But this work has already been useful even in its inefficient state, because it serves as a pretty definitive validation of our ideas about the function of imprinting in embryonic development, as well as the critical role methylation plays in this process. If we weren’t largely right about both of those, the efficiency of this approach wouldn’t be low—it would be zero.

PNAS, 2025. DOI: 10.1073/pnas.2425307122  (About DOIs).

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