Fig. 4: Conditional knockout of an essential gene using the Ribo-Off system. | Communications Biology

Fig. 4: Conditional knockout of an essential gene using the Ribo-Off system.

From: Ribo-On and Ribo-Off tools using a self-cleaving ribozyme allow manipulation of endogenous gene expression in C. elegans

Fig. 4: Conditional knockout of an essential gene using the Ribo-Off system.The alternative text for this image may have been generated using AI.

a Schematic illustration of using CRISPR/Cas9-mediated homologous recombination to insert the GFP::loxP::TTS::loxP::Rz cassette to the C-terminal of the endogenous bicd-1 locus, and the related outcome of protein expression with or without Cre expression. b Confocal fluorescent images to show the morphology of PVD dendrites. Cre recombinase was specifically expressed to delete the let-858 transcriptional terminator in various tissues using different tissue-specific promoters. L4-stage animals with the heat-shock::Cre transgene were treated with heat-shock. 3-day-old adult worms were imaged for all groups. Scale bars: 20 μm. c Quantifications of number of ectopic dendritic branches between the primary dendrites and the tertiary dendrites in a region 100 µm anterior to the PVD cell body in different genetic backgrounds as shown in (b). All values are presented as mean ± s.e.m. Number of animals quantified: n = 27 for WT; n = 20 for bicd-1(zac51); n = 26 for bicd-1::loxP::TTS::loxP::Rz without Cre; n = 26 for bicd-1::loxP::TTS::loxP::Rz with skin Cre; n = 28 for bicd-1::loxP::TTS::loxP::Rz with PVD Cre; n = 27 for bicd-1::loxP::TTS::loxP::Rz with muscle Cre. ns: not significant. ****p < 0.0001 (one-way ANOVA with the Tukey correction). Source data are provided as a Source Data file.

Back to article page