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. 2018 Oct 1;210(4):1355–1367. doi: 10.1534/genetics.118.301631

Figure 6.

Figure 6

An endogenous clustered regularly interspaced short palindromic repeats (CRISPR)-generated reporter verifies requirements for col-141 expression. (A) Utilizing CRISPR-Cas9 genome editing to knock out and replace col-141 with 2xNLS::gfp, creating a loss-of-function mutant and transcriptional reporter simultaneously. (B–D) GFP expression in col-141(lf) mutant. Dotted lines outline the approximate positions of the seam cells. Black arrow denotes a seam cell nucleus lacking GFP expression. (E–G) Loss of GFP expression in the sma-3 mutant background. (B and E) Fluorescent images; (C and F) Nomarski images; and (D and G) merged images. (H) Regulation of cuticle collagen gene expression by sma-3 in adult stages determined by quantitative reverse transcriptase PCR mirrors the transcriptional reporter. Results show the mean from four independent biological replicates with two technical replicates for each sample. Statistical analysis was performed by student’s t-test. ** P < 0.005 and *** P < 0.0005. WT, wild-type.