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Fig. 4 | Genome Biology

Fig. 4

From: Developmental dynamics of gene expression and alternative polyadenylation in the Caenorhabditis elegans germline

Fig. 4

In vivo expression of alternative 3′UTR isoforms in male and female germline. a Two-color in vivo reporter system to monitor the influence of different 3′UTR isoforms on protein expression. Fluorescent mCherry and GFP reporters are driven by the same promoter using an operon construct; co-transcriptional processing produces two trans-spliced (SL, splice leader) mRNAs carrying different 3′UTR isoforms. In the long isoform, the upstream PAS site is mutated to ensure usage of the downstream CPA site. b LITE-seq evidence for differential alternative 3′UTR isoform usage in vivo. Normalized LITE-Seq read counts for air-2 CPA sites in different samples. Representative CPA sites defining alternative 3′UTR isoforms are indicated on the gene model below (black triangles). A different 3′UTR isoform is preferred in female (long) and male (short) germline. The short isoform in the male is expressed at around twice the level of the long isoform in the female. c Alternative 3′UTR isoforms direct different patterns of protein expression in the female germline. Expression of the mCherry reporter, regulated by the short air-2 isoform, is similar to controls; GFP expression, regulated by the long isoform, is repressed in pachytene and oocytes relative to controls and reappears robustly in embryos. The distal tip of the germline, where cells begin mitotic proliferation, is marked with an asterisk

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