Figure 1.
Robust, tissue-specific labeling of a target protein at its native expression level. (A) NATF. CRISPR/Cas9-mediated gene editing is used to label a protein of interest with seven copies of GFP11 (GFP11x7). Transgenic expression of GFP1-10 with cell-specific promoters results in a bright, stable NATF fluorescent signal from multiple, reconstituted GFP molecules in specific tissues. (B) NATF workflow. Worms are injected with sgRNA, repair template, and co-injection markers. gfp11x7 knock-in worms are recovered after heat shock-induced excision of positive-selection genes. Crossing the gfp11x7 knock-in with gfp1-10 reporter lines results in tissue-specific labeling of the target protein with a reconstituted NATF-GFP tag. CRISPR, clustered regularly interspaced short palindromic repeats; GABA, γ-aminobutyric acid; NATF, Native And Tissue-specific Fluorescence; sgRNA, single-guide RNA.