Fig 1.
Schematic representation of the experimental design to evaluate the effectiveness of zygote cryopreservation compared with fresh zygotes to be used for CRISPR/Cas9 microinjection.
Table 1.
Experiment 1. Embryo survival, cleavage, development and mutation rate of CRISPR/Cas9 microinjected fresh or vitrified/warmed B6D2F1/J hybrid zygotes.
Fig 2.
Representative DIC images of in vitro developed embryos (Experiment 1).
Microinjection of fresh zygotes (A) or after vitrification-warming (D) resulted in two-cell embryos and expanded blastocysts upon incubation (B and C injected as fresh; E and F injected after vitrification-warming).
Fig 3.
DNA samples were obtained from blastocysts that were microinjected in zygote stage. Samples 0 and 2 were positive for indels; samples 1 and 3 were negative for indels. WT = wild-type blastocyst; NTC = PCR negative control. Arrows indicate heterodimers. Asterisk indicate the wild-type allele. 1 kb plus DNA ladder (Thermofisher) was used.
Table 2.
Experiment 2. Embryo survival, cleavage, pregnancy, birth and mutation rates of CRISPR/Cas9 microinjected fresh or vitrified/warmed C57BL/6J zygotes.