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Fig. 3

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Publication: Liu et al., 2022 - Cre/lox regulated conditional rescue and inactivation with zebrafish UFlip alleles generated by CRISPR-Cas9 targeted integration
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Fig. 3

Dre mRNA injection into rbbp4^off^/+ embryos leads to inversion of the UFlip cassette and efficient germline recovery of an inverted rbbp4^on allele.

(A) Diagram illustrating Dre-mediated inversion of the rbbp4^off allele to the on orientation. Repeated inversion of the cassette will continue as long as Dre is present. The final allele is predicted to be in the inverted ‘on’ orientation at a frequency of 50%. (B) PCR junction analysis of 8 embryos from an F1 adult that had been injected with Dre mRNA at the one-cell stage. Three embryos positive for expression of the lens BFP secondary marker show the expected 5’ and 3’ junction PCR amplicons for the inverted rbbp4^on allele. (C) Sequence analysis confirms Dre-mediated inversion of the cassette from the ‘off’ to ‘on’ orientation in BRP+/RFP - embryos.

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