ZFIN Image: McGrail et al., 2011, Fig. 3

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Figure Caption

Fig. 3 Two strategies for T2/OncZ insertional mutagenesis in zebrafish somatic tissues.

(A) Methods 1: Injection of in vitro transcribed SB11 mRNA at the 1-cell stage. (B) Method 2: Genetic cross between Tg(T2/OncZ, β-actin:RFP) (T2/OncZ) and constitutive Tg(Tol2<β-actin:SB11, cmlc2:GFP>) (β-actin:SB11) fish. At 24 hpf embryos are sorted into 4 progeny classes. (C) Excision PCR assay on 5 dpf larvae to demonstrate mobilization of transposons out of the concatemer in the presence of SB11 transposase. Primers 1 and 4 amplify a 220 bp band (red asterisk) flanking the transposon excision site in the concatemer. Left panel, Tg(T2/OncZ, β-actin:RFP)^is6 larvae; middle panel, SB11 injected Tg(T2/OncZ, β-actin:RFP)^is6 larvae; right panel, double transgenic Tg(T2/OncZ, β-actin:RFP)^is6; Tg(Tol2<β-actin:SB11, cmlc2:GFP>) larvae.

Acknowledgments

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