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F1 hybrid progenies between related subspecies often show hybrid sterility (HS) or inviability. HS is caused by failure of meiotic chromosome synapsis and sex body formation in house mouse. Previous studies identified two HS critical genomic regions named  Hstx2  on Chr X and  Hst 1 on Chr 17 by murine forward genetic approaches. HS gene on  Hst1  was reported to be  Prdm9 . Intersubspecific polymorphisms of  Prdm9  induce HS in hybrids, and  Prdm9  null mutation leads to sterility in the inbred strain. However, HS gene on  Hstx 2 remains unknown. Here, using knock-out studies, we showed that HS candidate genes on  Hstx2  are not individually essential for spermatogenesis in B6 strain. We examined 12 genes on  Hstx2 :  Ctag2 ,  49 3 0 4 47F04Rik ,  Mir743 ,  Mir46 5 d ,  Mir4 6 5c-2 ,  Mir465b-1 ,  Mir465c-1 ,  Mir465 ,  Gm1140 ,  Gm14692 ,  4933436I01Rik , and  Gm6812 . These genes were expressed in adult testes, and showed intersubspecific polymorphisms on expressed regions. This first reverse genetic approach to identify HS gene on  Hstx2  suggested that the loss of function of any one HS candidate gene does not cause complete sterility, unlike  Prdm9 . Thus, the mechanism(s) of HS by the HS gene on  Hstx2  might be different from that of  Prdm9 .

Reverse genetics reveals single gene of every candidate on Hybrid sterility, X Chromosome QTL 2 (Hstx2) are dispensable for spermatogenesis