FIGURE SUMMARY
Title

Generation of a conditional lima1a allele in zebrafish using the FLEx switch technology

Authors
Jungke, P., Hans, S., Gupta, M., Machate, A., Zöller, D., Brand, M.
Source
Full text @ Genesis
Fig. 2

Tg(FLEx-lima1a:GFP) recapitulates endogenous lima1a expression. (A) Schematic representation of SAGFLEx integrated into intron 2 of the lima1a locus in Tg(FLEx-lima1a:GFP). Native GFP fluorescence (B–F) and gfp transcription (G–K) in Tg(FLEx-lima1a:GFP) compared to lima1a expression in wildtype embryos (L–P). Lateral views at 12 somite stage (15 hpf), 20 somite stage (19 hpf), 24, 30, and 48 hpf. Dorsal views of single and double in situ hybridizations for lima1a and gfp at 30 hpf (Q–S).

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: 10-13 somites to Long-pec
Fig. 3

Functional consequences of the integration of SAGFLEx into the lima1a locus. (A) Schematic representation of SAGFLEx integrated into intron 2 of the lima1a locus in Tg(FLEx-lima1a:GFP). The positions of the primers used for qRT-PCR analysis (GFP-for/GFP-rev, E9-for/E9-rev) and the lima1a exon 9 probe are indicated. (B) Mating of heterozygous Tg(FLEx-lima1a:GFP) animals results in offspring with no, medium and strong GFP fluorescence. (C–E) Expression of lima1a in embryos with no, medium and strong GFP fluorescence (no GFP, GFP+, GFP++) using an exon 9 probe. Lateral and dorsal views at 28 hpf. (F) qRT-PCR analysis of lima1a exon 9 expression in embryos with medium and strong GFP fluorescence compared to siblings with no GFP fluorescence. ***P value ≥ 0,001. (G, H) Cellular localization of GFP fusions with wildtype Lima1a encoded by exons 1–9 (E1–E9) and truncated Lima1a encoded by exons 1–2 (E1–E2).
Fig. 4

Flp-mediated inversion and excision of SAGFLEx. (A) Schematic representation of the events of SAGFLEx integrated into intron 2 of the lima1a locus in Tg(FLEx-lima1a:GFP) after Flp mRNA injection. The position of the primers used for the allele-specific PCR (E2-for/pA-for/GFP-rev) are indicated. (B) Native GFP fluorescence in uninjected and Flp mRNA injected Tg(FLEx-lima1a:GFP) embryos. Lateral and dorsal views at 28 hpf. (C) Allele-specific PCR of uninjected and Flp mRNA injected Tg(FLEx-lima1a:GFP) embryos with the primers indicated above.
Fig. 5

Cre-mediated re-inversion of SAGFLEx. (A) Schematic representation of the events of SAGFLEx integrated into intron 2 of the lima1a locus in Tg(FLEx-lima1a:GFP) after Cre expression. The positions of the primers used for the allele-specific PCR (E2-for/pA-for/GFP-rev) are indicated. (B) Expression of Cre in rhombomeres 3 and 5 (r3/5) of the developing hindbrain results in the restoration of native GFP fluorescence. (C) Allele-specific PCR of GFP-positive and negative Tg(FLEx-lima1:GFP)inv embryos with the primers indicated above.
Fig. S1

[n/a]

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Terms:
Stage Range: 5-9 somites to Prim-15
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Genesis
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