Objective In order to evaluate the value of open pulled straw(OPS) method on cryopreservation of embryos of different genotypes of HBV transgenic mice, and to provide a basis for establishment of transgenic embryos bank. Methods Ethylene glygol (EG) was used as the cryoprotectant for cryopreservation of morula and blastocyst embryos of P 21 / , P 21 HBsAg/HBsAg and P 21 HBx/HBx . Result The average numbers of embryo production of P 21 / , P 21 HBsAg/HBsAg and P 21 HBx/HBx were 27 89, 15 56 and 9 14, respectively, after superovulatory treatment. The recovery rates of P 21 / , P 21 HBsAg/HBsAg and P 21 HBx/HBx were 88 7%, 81 1% and 80 3%,respectively. The development rates after thawing were 82 6%, 79 4% and 56 6% in vitro, respectively. The results indicated that the numbers of superovulation of P 21 HBsAg/HBsAg and P 21 HBx/HBx were lower than those of P 21 / . The recovery rates were not significantly different in the three genotypes mice. The survival rate after thawing of P 21 HBx/HBx was lower than that of P 21 / and P 21 HBsAg/HBsAg . Conclusion The genotype of mice affect the results of superovulation number, recovery rate and survival of embryos. The OPS method is feasible and useful in cryopreservation of transgenic mouse embryos.