{"doi":"10.1111/pbi.13010","title":"Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters","abstract":"<jats:title>Summary</jats:title><jats:p><jats:italic>Cis</jats:italic>‐regulatory elements in promoters are major determinants of binding specificity of transcription factors (<jats:styled-content style=\"fixed-case\">TF</jats:styled-content>s) for transcriptional regulation. To improve our understanding of how these short <jats:styled-content style=\"fixed-case\">DNA</jats:styled-content> sequences regulate gene expression, synthetic promoters consisting of both classical (<jats:styled-content style=\"fixed-case\">CACGTG</jats:styled-content>) and variant G‐box core sequences along with different flanking sequences derived from the promoters of three different highly expressing soybean genes, were constructed and used to regulate a <jats:italic>green fluorescent protein</jats:italic> (<jats:italic>gfp</jats:italic>) gene. Use of the classical 6‐bp G‐box provided information on the base level of <jats:styled-content style=\"fixed-case\">GFP</jats:styled-content> expression while modifications to the 2–4 flanking bases on either side of the G‐box influenced the intensity of gene expression in both transiently transformed lima bean cotyledons and stably transformed soybean hairy roots. The proximal 2‐bp sequences on either flank of the G‐box significantly affected G‐box activity, while the distal 2‐bp flanking nucleotides also influenced gene expression albeit with a decreasing effect. Manipulation of the upstream 2‐ to 4‐bp flanking sequence of a G‐box variant (<jats:styled-content style=\"fixed-case\">GACGTG</jats:styled-content>), found in the proximal region of a relatively weak soybean glycinin promoter, significantly enhanced promoter activity using both transient and stable expression assays, if the G‐box variant was first converted into a classical G‐box (<jats:styled-content style=\"fixed-case\">CACGTG</jats:styled-content>). In addition to increasing our understanding of regulatory element composition and structure, this study shows that minimal targeted changes in native promoter sequences can lead to enhanced gene expression, and suggests that genome editing of the promoter region can result in useful and predictable changes in native gene expression.</jats:p>","journal":"Plant Biotechnology Journal","year":2019,"id":35328,"datarank":1.0887776072970263,"base_score":3.367295829986474,"endowment":3.367295829986474,"self_citation_contribution":0.5050943744979712,"citation_network_contribution":0.5836832327990552,"self_endowment_contribution":0.5050943744979712,"citer_contribution":0.5836832327990552,"corpus_percentile":null,"corpus_rank":null,"citation_count":28,"citer_count":28,"citers_with_citation_signal":24,"citers_with_endowment":24,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":181168,"name":"Leah K. McHale","orcid":null,"position":1,"is_corresponding":false},{"id":181169,"name":"John J. Finer","orcid":"0000-0001-8004-5468","position":2,"is_corresponding":false},{"id":29979,"name":"Ning Zhang","orcid":"0000-0001-6643-6980","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.367295829986474,"endowment":3.367295829986474,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"30191675","pmcid":"PMC6419578","openalex_id":"https://openalex.org/W2889605195","authors":[],"funders":[{"funder_name":"United Soybean Board","grant_id":"","title":null},{"funder_name":"Ohio Agricultural Research and Development Center, Ohio State University","grant_id":"","title":null}],"total_grants":2,"fwci":0.9276,"citation_percentile":0.74590933,"influential_citations":0,"citation_trend":[{"year":2019,"count":4},{"year":2020,"count":3},{"year":2021,"count":3},{"year":2022,"count":5},{"year":2023,"count":1},{"year":2024,"count":7},{"year":2025,"count":3},{"year":2026,"count":2}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/pbi.13010","host_type":"journal"},{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/pbi.13010","host_type":"GOLD"},{"url":"https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/pbi.13010","host_type":"publisher"},{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fpbi.13010","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1111/pbi.13010","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pbi.13010","host_type":"publisher"},{"url":"https://doi.org/10.1111/pbi.13010","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/30191675","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/6419578","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC6419578","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC6419578?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["CRISPR and Genetic Engineering","Plant Virus Research Studies","Plant tissue culture and regeneration","Biology","Medicine","Environmental Science","Cotyledon","Gene Expression Regulation, Plant","Genes, Reporter","Globulins","Phaseolus","Plant Roots","Plants, Genetically Modified","Promoter Regions, Genetic","Soybean Proteins","Glycine max","Transcription Factors"],"mesh_terms":["Globulins","Promoter Regions, Genetic","Glycine max","Transcription Factors","Genes, Reporter","Gene Expression Regulation, Plant","Plant Roots","Cotyledon","Phaseolus","Soybean Proteins","Plants, Genetically Modified"],"keywords":["Promoter","Biology","Gene","Regulatory sequence","5' flanking region","CAAT box","Gene expression","Genetics","TATA box","Transcription (linguistics)","Regulation of gene expression","Transcription factor","DNA","Molecular biology","Flanking sequences","cis-regulatory element","G-box","Synthetic Promoter","Glycinin Promoter","Elongation Factor-1 Α Promoter"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life in Land"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-10T03:12:15.960683Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}