biostudies-literatureGorjifard SUnited States Department of Agriculture | National Institute of Food and Agriculture (NIFA)NHGRI NIH HHSNational Science Foundation (NSF)U.S. Department of Health & Human Services | National Institutes of Health (NIH)U.S. Department of Health & Human Services | National Institutes of HealthUnited States Department of Agriculture | National Institute of Food and AgricultureNIGMS NIH HHSNational Science Foundation5868https://www.ebi.ac.uk/biostudies/studies/S-EPMC11245536biostudies-literatureUnknown15(1)The 3' end of a gene, often called a terminator, modulates mRNA stability, localization, translation, and polyadenylation. Here, we adapted Plant STARR-seq, a massively parallel reporter assay, to measure the activity of over 50,000 terminators from the plants Arabidopsis thaliana and Zea mays. We characterize thousands of plant terminators, including many that outperform bacterial terminators commonly used in plants. Terminator activity is species-specific, differing in tobacco leaf and maize protoplast assays. While recapitulating known biology, our results reveal the relative contributions of polyadenylation motifs to terminator strength. We built a computational model to predict terminator strength and used it to conduct in silico evolution that generated optimized synthetic terminatorNature communicationsArabidopsis and maize terminator strength is determined by GC content, polyadenylation motifs and cleavage probability.PMC112455362023-67012-394452240888HG000035R01-GM079712R01 GM079712T32 HG000035R35 GM1395321R35GM139532Bubb KJores TQueitsch CBuckler ESTonnies JWrightsman TFields SCuperus JTGorjifard SMueth NAfalseArabidopsis and maize terminator strength is determined by GC content, polyadenylation motifs and cleavage probability.The 3' end of a gene, often called a terminator, modulates mRNA stability, localization, translation, and polyadenylation. Here, we adapted Plant STARR-seq, a massively parallel reporter assay, to measure the activity of over 50,000 terminators from the plants Arabidopsis thaliana and Zea mays. We characterize thousands of plant terminators, including many that outperform bacterial terminators commonly used in plants. Terminator activity is species-specific, differing in tobacco leaf and maize protoplast assays. While recapitulating known biology, our results reveal the relative contributions of polyadenylation motifs to terminator strength. We built a computational model to predict terminator strength and used it to conduct in silico evolution that generated optimized synthetic terminator2024-01-01T00:00:00Z2024 Jul2025-04-18T21:43:03.887Z2025-02-19T02:34:39.171ZS-EPMC112455363899725210.1038/s41467-024-50174-7