FIGURE

Figure 2.

ID

ZDB-FIG-230308-14

Publication

Mi et al., 2023 - Efficient knock-in method enabling lineage tracing in zebrafish

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Figure 2.

The design and characterization of knock-in lines at the <italic toggle='yes'>nkx6.1</italic> locus.

(A) The design of donor dsDNA template and gRNA sequence for the construction of knock-in lines at the 3′ end of the nkx6.1 locus. The nucleotide sequence in blue indicates gRNA; whereas the nucleotide sequence in red indicates the PAM sequence. The bottom panel shows the sequences of LHA and RHA. (B, C, D) Summary statistics of nkx6.1 knock-in efficiency, including the percentage of injected F0 with observable fluorescence labelling in the hindbrain and spinal cord (B), the percentage of adult F0 giving rise to germline transmission (out of those that bred) (C) and the percentage of F1 siblings inheriting each allele (D). (E, F, G) Representative confocal images of TgKI(nkx6.1-p2A-mNeonGreen-t2A-iCre);Tg(ubi:CSHm) at 1 (E), 2 (F), and 3 dpf (G). Cells expressing mNeonGreen indicate nkx6.1⁺ cells; all progenies of nkx6.1⁺ cells were labelled with H2BmCherry. The insets are magnified views showing the expression pattern of two fluorescent proteins. (H) Representative confocal image of lineage-tracing results in the principal islet of the pancreas in the TgKI(nkx6.1-p2A-mNeonGreen-t2A-iCre);Tg(ubi:CSHm) line at 6 dpf. Cells in white shown in H are β-cells with Insulin staining, whereas cells in green in H’’ are α-cells with Glucagon staining. (E, F, G, H) Scale bars = 200 μm (E, F, G) or 20 μm (H).

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