Figure 2. TALEN-mediated knockout of Cnot6l gene in mice.

(A) A scheme of Cnot6l gene depicting the position of the deletion in the genomic DNA and the corresponding part of the CNOT6L protein. Protein domains were mapped using the Conserved Domain Database search (43). (B) Sequences of two alleles identified in F₀ animals, which carry ∼31.3-kb deletions of exons 4–11. Underlined sequences indicate TALEN cognate sequences. A short fragment of mitochondrial DNA integrated into the deleted locus is visualized in lowercase red font. (C) Zygotes from in vitro–fertilized Cnot6l^−/− eggs develop significantly slower (t test, P-value < 0.001 for all stages) than zygotes developing from heterozygous eggs. Error bar = SD. In total, 95 and 133 zygotes produced from Cnot6l^−/− (−/−) and Cnot6l^+/− (+/−), respectively, were analyzed using the PrimoVision time-lapse system. Numbers indicate hours from the point the zygotes were placed into the tracking system (5 h after mixing sperm with cumulus oocyte complex), which automatically detects the first four cleavage events and formation of early blastocysts.