Fig. 4: The effect of viral 2A peptides and operon sequences on germline co-expression of genes.

a Germline expression of a ce-gfp fused to a genomic Psmu-1::smu-1 gene (germline and soma) separated by various viral 2A peptide tags. 2A tags result in ribosomal skipping and peptide cleavage, which allows co-expression of two or more genes from one open reading frame⁵⁴. n = 10, 9, 8, and 9 biologically independent transgenic lines (from top to bottom). b Germline expression of a ce-gfp fused to a genomic Ppie-1::smu-1 gene (germline-specific) separated by various 2 A peptide tags. n = 8, 8, 3, and 8 biologically independent transgenic lines (from top to bottom). c Germline expression of a ce-gfp fused to a genomic Psmu-1::smu-1 gene (germline and soma) separated by various operon sequences. Operons are common in C. elegans⁵⁵, particularly for genes expressed in the germline⁵⁶. Operons allow co-expression or more than one gene from a single promoter. n = 11, 5, 8, and 7 biologically independent transgenic lines (from top to bottom). d Germline expression of a ce-gfp fused to a genomic Ppie-1::smu-1 gene (germline and soma) separated by various operon sequences. n = 2, 6, 9, and 8 biologically independent transgenic lines (from top to bottom). Transgene scale bars = 100 nucleotides. All germline fluorescence was quantified from transgenic animals carrying simple extrachromosomal arrays imaged with a ×40 or ×63 oil objective at 25 °C. Each datapoint indicates one independent measurement of germline fluorescence scored from 11 animals from an independent transgenic line. Bars indicate the mean, and error bars indicate the SEM. Source data are available in the Source Data file.