{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T01:30:40Z","timestamp":1784770240490,"version":"3.55.0"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T00:00:00Z","timestamp":1754092800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T00:00:00Z","timestamp":1754092800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01 DK082412"],"award-info":[{"award-number":["R01 DK082412"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01 DK082412"],"award-info":[{"award-number":["R01 DK082412"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The digestive enzyme trypsin is an important driver of pancreatitis development and progression. Inhibition of intrapancreatic trypsin activity is an attractive yet so far unsuccessful approach for the treatment and prevention of pancreatitis. Gene therapy using adeno-associated viral vector containing the human serine protease inhibitor Kazal type 1 (\n                    <jats:italic>SPINK1<\/jats:italic>\n                    ) gene has shown efficacy against murine pancreatitis in preclinical experiments. To advance this method to clinical testing, viral vectors with improved inhibitor expression are needed. In this study, we investigated the effect of minigenes with single introns on the expression of the mouse\n                    <jats:italic>Spink1<\/jats:italic>\n                    gene. We found that minigenes markedly stimulated the expression of mouse\n                    <jats:italic>Spink1<\/jats:italic>\n                    mRNA and SPINK1 protein in HEK 293T cells transfected with plasmids and in AR42J rat pancreatic acinar cells and isolated mouse pancreatic acini transduced with adenoviral vectors. The optimal intron size for expression enhancement was 100 nucleotides. We also demonstrated that mouse\n                    <jats:italic>Spink1<\/jats:italic>\n                    mRNA expressed from a cDNA construct underwent spurious mRNA splicing and minigenes prevented this splicing defect. However, enhancement of mouse\n                    <jats:italic>Spink1<\/jats:italic>\n                    expression by minigenes was unrelated to the rescue of aberrant mRNA splicing. The observations will facilitate the development of advanced viral vectors harboring mouse\n                    <jats:italic>Spink1<\/jats:italic>\n                    for preclinical therapeutic studies in mouse models of pancreatitis.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-13128-7","type":"journal-article","created":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T07:13:52Z","timestamp":1754118832000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Minigenes enhance heterologous expression and prevent aberrant splicing of mouse Spink1"],"prefix":"10.1038","volume":"15","author":[{"given":"Gerg\u0151","family":"Berke","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikl\u00f3s","family":"Sahin-T\u00f3th","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,8,2]]},"reference":[{"key":"13128_CR1","doi-asserted-by":"publisher","first-page":"314","DOI":"10.1007\/BF01318124","volume":"31","author":"H Rinderknecht","year":"1986","unstructured":"Rinderknecht, H. Activation of pancreatic zymogens. Normal activation, premature intrapancreatic activation, protective mechanisms against inappropriate activation. Dig. Dis. Sci. 31, 314\u2013321 (1986).","journal-title":"Dig. Dis. Sci."},{"key":"13128_CR2","doi-asserted-by":"publisher","first-page":"1692","DOI":"10.1007\/s10620-017-4601-3","volume":"62","author":"E Hegyi","year":"2017","unstructured":"Hegyi, E. & Sahin-T\u00f3th, M. Genetic risk in chronic pancreatitis: the trypsin-dependent pathway. Dig. Dis. Sci. 62, 1692\u20131701 (2017).","journal-title":"Dig. Dis. Sci."},{"key":"13128_CR3","doi-asserted-by":"publisher","first-page":"100343","DOI":"10.1016\/j.jbc.2021.100343","volume":"296","author":"A Szab\u00f3","year":"2021","unstructured":"Szab\u00f3, A. et al. Defective binding of SPINK1 variants is an uncommon mechanism for impaired trypsin Inhibition in chronic pancreatitis. J. Biol. Chem. 296, 100343 (2021).","journal-title":"J. Biol. Chem."},{"key":"13128_CR4","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1016\/j.pan.2023.04.043","volume":"23","author":"NC Morales Granda","year":"2023","unstructured":"Morales Granda, N. C., Toldi, V., Miczi, M., Lassoued, M. & Szab\u00f3, A. Inhibition of mouse trypsin isoforms by SPINK1 and effect of human pancreatitis-associated mutations. Pancreatology 23, 358\u2013366 (2023).","journal-title":"Pancreatology"},{"key":"13128_CR5","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1038\/76088","volume":"25","author":"H Witt","year":"2000","unstructured":"Witt, H. et al. Mutations in the gene encoding the Serine protease inhibitor, Kazal type 1 are associated with chronic pancreatitis. Nat. Genet. 25, 213\u2013216 (2000).","journal-title":"Nat. Genet."},{"key":"13128_CR6","doi-asserted-by":"publisher","first-page":"696","DOI":"10.1016\/j.gastro.2005.05.057","volume":"129","author":"M Ohmuraya","year":"2005","unstructured":"Ohmuraya, M. et al. Autophagic cell death of pancreatic acinar cells in Serine protease inhibitor Kazal type 3-deficient mice. Gastroenterology 129, 696\u2013705 (2005).","journal-title":"Gastroenterology"},{"key":"13128_CR7","doi-asserted-by":"publisher","first-page":"G518","DOI":"10.1152\/ajpgi.00431.2009","volume":"298","author":"JM Romac","year":"2010","unstructured":"Romac, J. M. et al. Transgenic expression of pancreatic secretory trypsin inhibitor-1 rescues SPINK3-deficient mice and restores a normal pancreatic phenotype. Am. J. Physiol. Gastrointest. Liver Physiol. 298, G518\u2013524 (2010).","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"13128_CR8","doi-asserted-by":"publisher","first-page":"101361","DOI":"10.1016\/j.jcmgh.2024.05.009","volume":"18","author":"A Demcs\u00e1k","year":"2024","unstructured":"Demcs\u00e1k, A. & Sahin-T\u00f3th, M. Heterozygous Spink1 deficiency promotes trypsin-dependent chronic pancreatitis in mice. Cell. Mol. Gastroenterol. Hepatol. 18, 101361 (2024).","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"13128_CR9","doi-asserted-by":"publisher","first-page":"967","DOI":"10.1136\/gutjnl-2019-318790","volume":"69","author":"C Sun","year":"2020","unstructured":"Sun, C. et al. Heterozygous Spink1 c.194\u2009+\u20092T\u2009>\u2009C mutant mice spontaneously develop chronic pancreatitis. Gut 69, 967\u2013968 (2020).","journal-title":"Gut"},{"key":"13128_CR10","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1016\/j.pan.2024.05.514","volume":"24","author":"M Liu","year":"2024","unstructured":"Liu, M. et al. Heterozygous Spink1 c.194\u2009+\u20092T\u2009>\u2009C mutation promotes chronic pancreatitis after acute attack in mice. Pancreatology 24, 677\u2013689 (2024).","journal-title":"Pancreatology"},{"key":"13128_CR11","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1053\/j.gastro.2004.11.052","volume":"128","author":"JD Nathan","year":"2005","unstructured":"Nathan, J. D. et al. Transgenic expression of pancreatic secretory trypsin inhibitor-I ameliorates secretagogue-induced pancreatitis in mice. Gastroenterology 128, 717\u2013727 (2005).","journal-title":"Gastroenterology"},{"key":"13128_CR12","doi-asserted-by":"publisher","first-page":"e24","DOI":"10.1097\/MPA.0b013e3181bc45e9","volume":"39","author":"JD Nathan","year":"2010","unstructured":"Nathan, J. D. et al. Protection against chronic pancreatitis and pancreatic fibrosis in mice overexpressing pancreatic secretory trypsin inhibitor. Pancreas 39, e24\u201330 (2010).","journal-title":"Pancreas"},{"key":"13128_CR13","doi-asserted-by":"publisher","first-page":"G535","DOI":"10.1152\/ajpgi.00287.2011","volume":"302","author":"JM Romac","year":"2012","unstructured":"Romac, J. M. et al. Pancreatic secretory trypsin inhibitor I reduces the severity of chronic pancreatitis in mice overexpressing interleukin-1\u03b2 in the pancreas. Am. J. Physiol. Gastrointest. Liver Physiol. 302, G535\u2013541 (2012).","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"13128_CR14","doi-asserted-by":"publisher","first-page":"1142","DOI":"10.1136\/gutjnl-2023-330788","volume":"73","author":"YC Wang","year":"2024","unstructured":"Wang, Y. C. et al. Pancreas-directed AAV8-hSPINK1 gene therapy safely and effectively protects against pancreatitis in mice. Gut 73, 1142\u20131155 (2024).","journal-title":"Gut"},{"key":"13128_CR15","doi-asserted-by":"publisher","first-page":"545","DOI":"10.1136\/gut.2008.164947","volume":"58","author":"E Kereszturi","year":"2009","unstructured":"Kereszturi, E., Kir\u00e1ly, O. & Sahin-T\u00f3th, M. Minigene analysis of intronic variants in common SPINK1 haplotypes associated with chronic pancreatitis. Gut 58, 545\u2013549 (2009).","journal-title":"Gut"},{"key":"13128_CR16","doi-asserted-by":"publisher","first-page":"e9","DOI":"10.1136\/gutjnl-2024-332818","volume":"74","author":"G Berke","year":"2025","unstructured":"Berke, G. & Sahin-T\u00f3th, M. Intron-mediated enhancement of SPINK1 expression for pancreatitis therapy. Gut 74, e9 (2025).","journal-title":"Gut"},{"key":"13128_CR17","doi-asserted-by":"publisher","first-page":"342","DOI":"10.1016\/j.pan.2021.01.007","volume":"21","author":"D Mosztbacher","year":"2021","unstructured":"Mosztbacher, D. & Sahin-T\u00f3th, M. Mouse model suggests limited role for human mesotrypsin in pancreatitis. Pancreatology 21, 342\u2013352 (2021).","journal-title":"Pancreatology"},{"key":"13128_CR18","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1136\/gut.2009.198903","volume":"59","author":"R Szmola","year":"2010","unstructured":"Szmola, R. & Sahin-T\u00f3th, M. Pancreatitis-associated chymotrypsinogen C (CTRC) mutant elicits endoplasmic reticulum stress in pancreatic acinar cells. Gut 59, 365\u2013372 (2010).","journal-title":"Gut"},{"key":"13128_CR19","doi-asserted-by":"publisher","DOI":"10.3998\/panc.2010.18","author":"JA Williams","year":"2010","unstructured":"Williams, J. A. Isolation of rodent pancreatic acinar cells and acini by collagenase digestion. Pancreapedia: Exocrine Pancreas Knowl. Base. https:\/\/doi.org\/10.3998\/panc.2010.18 (2010).","journal-title":"Pancreapedia: Exocrine Pancreas Knowl. Base"},{"key":"13128_CR20","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1136\/gutjnl-2018-315994","volume":"68","author":"E Hegyi","year":"2019","unstructured":"Hegyi, E. & Sahin-T\u00f3th, M. Human CPA1 mutation causes digestive enzyme misfolding and chronic pancreatitis in mice. Gut 68, 301\u2013312 (2019).","journal-title":"Gut"},{"key":"13128_CR21","doi-asserted-by":"publisher","first-page":"1083","DOI":"10.1053\/j.gastro.2019.11.020","volume":"158","author":"Z Jancs\u00f3","year":"2020","unstructured":"Jancs\u00f3, Z. & Sahin-T\u00f3th, M. Mutation that promotes activation of trypsinogen increases severity of secretagogue-induced pancreatitis in mice. Gastroenterology 158, 1083\u20131094 (2020).","journal-title":"Gastroenterology"},{"key":"13128_CR22","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1016\/S0304-3835(79)80080-4","volume":"7","author":"DS Longnecker","year":"1979","unstructured":"Longnecker, D. S. et al. Transplantation of azaserine-induced carcinomas of pancreas in rats. Cancer Lett. 7, 197\u2013202 (1979).","journal-title":"Cancer Lett."},{"key":"13128_CR23","first-page":"212","volume":"16","author":"NW Jessop","year":"1980","unstructured":"Jessop, N. W. & Hay, R. J. Characteristics of two rat pancreatic exocrine cell lines derived from transplantable tumors. Vitro 16, 212 (1980). (Abstracts from the Thirty-First Annual Meeting of the Tissue Culture Association).","journal-title":"Vitro"},{"key":"13128_CR24","doi-asserted-by":"publisher","first-page":"1200","DOI":"10.1083\/jcb.100.4.1200","volume":"100","author":"CD Logsdon","year":"1985","unstructured":"Logsdon, C. D. et al. Glucocorticoids increase amylase mRNA levels, secretory organelles, and secretion in pancreatic acinar AR42J cells. J. Cell. Biol. 100, 1200\u20131208 (1985).","journal-title":"J. Cell. Biol."},{"key":"13128_CR25","first-page":"101","volume":"47","author":"B Swarovsky","year":"1988","unstructured":"Swarovsky, B. et al. Coupled induction of exocrine proteins and intracellular compartments involved in the secretory pathway in AR4-2J cells by glucocorticoids. Eur. J. Cell. Biol. 47, 101\u2013111 (1988).","journal-title":"Eur. J. Cell. Biol."},{"issue":"Pt B","key":"13128_CR26","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1016\/j.biocel.2017.06.016","volume":"91","author":"O Shaul","year":"2017","unstructured":"Shaul, O. How introns enhance gene expression. Int. J. Biochem. Cell. Biol. 91 (Pt B), 145\u2013155 (2017).","journal-title":"Int. J. Biochem. Cell. Biol."},{"key":"13128_CR27","doi-asserted-by":"publisher","first-page":"672","DOI":"10.3389\/fgene.2018.00672","volume":"9","author":"AB Rose","year":"2019","unstructured":"Rose, A. B. Introns as gene regulators: A brick on the accelerator. Front. Genet. 9, 672 (2019).","journal-title":"Front. Genet."},{"key":"13128_CR28","doi-asserted-by":"publisher","first-page":"618","DOI":"10.1261\/rna.5260303","volume":"9","author":"S Lu","year":"2003","unstructured":"Lu, S. & Cullen, B. R. Analysis of the stimulatory effect of splicing on mRNA production and utilization in mammalian cells. RNA 9, 618\u2013630 (2003).","journal-title":"RNA"},{"key":"13128_CR29","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1261\/rna.5250403","volume":"9","author":"A Nott","year":"2003","unstructured":"Nott, A., Meislin, S. H. & Moore, M. J. A quantitative analysis of intron effects on mammalian gene expression. RNA 9, 607\u2013617 (2003).","journal-title":"RNA"},{"key":"13128_CR30","doi-asserted-by":"publisher","first-page":"8505","DOI":"10.1073\/pnas.1112400109","volume":"109","author":"AM Moabbi","year":"2012","unstructured":"Moabbi, A. M., Agarwal, N., El Kaderi, B. & Ansari, A. Role for gene looping in intron-mediated enhancement of transcription. Proc. Natl. Acad. Sci. USA. 109, 8505\u20138510 (2012).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"13128_CR31","doi-asserted-by":"publisher","first-page":"11327","DOI":"10.1073\/pnas.1934877100","volume":"100","author":"HL Wiegand","year":"2003","unstructured":"Wiegand, H. L., Lu, S. & Cullen, B. R. Exon junction complexes mediate the enhancing effect of splicing on mRNA expression. Proc. Natl. Acad. Sci. USA. 100, 11327\u201311332 (2003).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"13128_CR32","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1016\/j.molcel.2018.08.030","volume":"72","author":"V Boehm","year":"2018","unstructured":"Boehm, V. et al. Exon junction complexes suppress spurious splice sites to safeguard transcriptome integrity. Mol. Cell. 72, 482\u2013495 (2018).","journal-title":"Mol. Cell."},{"key":"13128_CR33","doi-asserted-by":"publisher","first-page":"e1009563","DOI":"10.1371\/journal.pgen.1009563","volume":"17","author":"B Joseph","year":"2021","unstructured":"Joseph, B. & Lai, E. C. The exon junction complex and intron removal prevent re-splicing of mRNA. PLoS Genet. 17, e1009563 (2021).","journal-title":"PLoS Genet."},{"key":"13128_CR34","doi-asserted-by":"publisher","first-page":"210","DOI":"10.1101\/gad.1163204","volume":"18","author":"A Nott","year":"2004","unstructured":"Nott, A., Le Hir, H. & Moore, M. J. Splicing enhances translation in mammalian cells: an additional function of the exon junction complex. Genes Dev. 18, 210\u2013222 (2004).","journal-title":"Genes Dev."},{"key":"13128_CR35","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1016\/j.gde.2011.04.001","volume":"21","author":"CJ McManus","year":"2011","unstructured":"McManus, C. J. & Graveley, B. R. RNA structure and the mechanisms of alternative splicing. Curr. Opin. Genet. Dev. 21, 373\u2013379 (2011).","journal-title":"Curr. Opin. Genet. Dev."},{"key":"13128_CR36","doi-asserted-by":"publisher","first-page":"495","DOI":"10.1093\/dnares\/dsv028","volume":"22","author":"A Piovesan","year":"2015","unstructured":"Piovesan, A. et al. Identification of minimal eukaryotic introns through genebase, a user-friendly tool for parsing the NCBI gene databank. DNA Res. 22, 495\u2013503 (2015).","journal-title":"DNA Res."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-13128-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-13128-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-13128-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T13:36:14Z","timestamp":1761658574000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-13128-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,2]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["13128"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-13128-7","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,2]]},"assertion":[{"value":"28 May 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 July 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 August 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"28222"}}