Testis electroporation coupled with autophagy inhibitor to treat non-obstructive azoospermia

Infertility affects 10%-20% of the population in most coun-tries, with approximately half of those cases resulting from male infertility. Although millions of infertile men are able to have children with the assistance of reproductive technology, individuals with non-obstructive azoospermia (NOA) syn-drome are unable to do so because they lack functional sperm. Therefore, some other strategies for infertile NOA men are still urgently needed. Our current study uses an NOA-like mouse model to optimize microinjection and a subsequent electropo-ration method to test potential treatment strategies. We showed that the spermatogenetic process could be partially rescued in young Stra8-Rnf20-/- mice with microinjection and subsequent electroporation of Rnf20 plasmids into the testes. All meiotic prophase I stages could be identified, and programmed DNA double-strand break repair factors could successfully be recruited to Stra8-Rnf20-/- spermatocytes after electroporation. Moreover, by including an autophagy inhibi-tor in the treatment, electroporation significantly improved the spermatogenetic rescue efficiency of adult Stra8-Rnf20-/- mice. Most importantly, infertility caused by Rnf20 depletions could be overcome by electroporation coupled with intracyto-plasmic sperm injection. Our studies establish a relative safe and efficient testis electroporation system and provide a prom-ising therapeutic strategy for patients with NOA.