Extended Data Fig. 6: Cell-specific knockdown system is efficient to disrupt genes in oligodendrocytes in zebrafish. | Nature Neuroscience

Extended Data Fig. 6: Cell-specific knockdown system is efficient to disrupt genes in oligodendrocytes in zebrafish.

From: Synaptic input and Ca2+ activity in zebrafish oligodendrocyte precursor cells contribute to myelin sheath formation

Extended Data Fig. 6

(a) Schematic of the plasmid (10xUAS:myrmScarlet-p2A-Cas9, U6:sgRNA1;U6:sgRNA2) used to induce cell-specific Cas9 expression with membrane labeling and 2 separate U6-driven sgRNAs. (b) Representative gel images of digested PCR product of genomic regions targeted by the sgRNAs. Each lane represents a single embryo at 1 dpf. The left 4 lanes are uninjected (-) and the right 4 lanes are injected (+) with sgRNAs and Cas9 protein. (c) Schematic of larva with sparsely labeled oligodendrocyte lineage cells resulting from Tg(sox10:Kalta4) crossed with transgenic fish carrying the plasmid from (a). (d) Representative images of single oligodendrocytes in the spinal cord of Tg(10xUAS:myrmScarlet-p2A-Cas9, U6:ctrl-sgRNA1;U6:ctrl-sgRNA2) and Tg(10xUAS:myrmScarlet-p2A-Cas9, U6:myrfex3-sgRNA;U6:myrfex4-sgRNA) at 6 dpf. Knockdown of Myrf serves as a control for our targeting approach. (e) Average myelin sheath length in (d). n = 24 and 22 cells from 15 fish; t[44] = 2.247. All data are represented as mean ± SEM; (e) two-sided unpaired t-test; scale bars 5 μm.

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