{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T15:01:30Z","timestamp":1761663690923,"version":"3.41.0"},"reference-count":39,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2018,2,1]],"date-time":"2018-02-01T00:00:00Z","timestamp":1517443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,2,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Von Willebrand Disease (VWD) type 3 is a serious and sometimes fatal hereditary bleeding disorder. In pigs, the disease has been known for decades, and affected animals are used as models for the human disease. Due to the recessive mode of inheritance of VWD type 3, severe bleeding is typically seen in homozygous individuals. We sequenced the complete porcine VWF (Von Willebrand Factor) complementary DNA (cDNA) and detected a tandem duplication of exons 17 and 18, causing a frameshift and a premature termination codon (p.Val814LeufsTer3) in the affected pig. Subsequent next generation sequencing on genomic DNA proved the existence of a 12.3-kb tandem duplication associated with VWD. This duplication putatively originates from porcine Short Interspersed Nuclear Elements (SINEs) located within VWF introns 16 and 18 with high identity. The premature termination truncates the VWF open reading frame by a large part, resulting in an almost entire loss of the mature peptide. It is therefore supposed to account for the severe VWD type 3. Our results further indicate the presence of strong, nonsense-mediated decay in VWF messenger RNA (mRNA) containing the duplication, which was supported by the almost complete absence of the complete VWF protein in immunohistochemistry analysis of the VWD-affected pig. In the past, differentiation of wild-type and heterozygous pigs in this VWD colony had to rely on clinical examinations and additional laboratory methods. The present study provides the basis to distinguish both genotypes by performing a rapid and simple genetic analysis.<\/jats:p>","DOI":"10.1534\/g3.117.300432","type":"journal-article","created":{"date-parts":[[2017,12,6]],"date-time":"2017-12-06T01:45:23Z","timestamp":1512524723000},"page":"577-585","source":"Crossref","is-referenced-by-count":5,"title":["A 12.3-kb Duplication Within the<i>VWF<\/i>Gene in Pigs Affected by Von Willebrand Disease Type 3"],"prefix":"10.1093","volume":"8","author":[{"given":"Stefanie","family":"Lehner","sequence":"first","affiliation":[{"name":"Department of Fundamental Research, Werlhof Institute GmbH, 30159 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mahnaz","family":"Ekhlasi-Hundrieser","sequence":"additional","affiliation":[{"name":"Department of Fundamental Research, Werlhof Institute GmbH, 30159 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carsten","family":"Detering","sequence":"additional","affiliation":[{"name":"Department of Fundamental Research, Werlhof Institute GmbH, 30159 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanna","family":"Allerkamp","sequence":"additional","affiliation":[{"name":"Department of Fundamental Research, Werlhof Institute GmbH, 30159 Hannover, Germany"},{"name":"Institute for Anatomy, University of Veterinary Medicine Hannover Foundation, 30173 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christiane","family":"Pfarrer","sequence":"additional","affiliation":[{"name":"Institute for Anatomy, University of Veterinary Medicine Hannover Foundation, 30173 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mario","family":"von Depka Prondzinski","sequence":"additional","affiliation":[{"name":"Department of Fundamental Research, Werlhof Institute GmbH, 30159 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2018,2,1]]},"reference":[{"key":"2021042013282762400_bib1","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nmeth0410-248","article-title":"A method and server for predicting damaging missense mutations.","volume":"7","author":"Adzhubei","year":"2010","journal-title":"Nat. Methods"},{"key":"2021042013282762400_bib2","doi-asserted-by":"crossref","first-page":"W3","DOI":"10.1093\/nar\/gkw343","article-title":"The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update.","volume":"44","author":"Afgan","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"2021042013282762400_bib3","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1182\/blood.V72.1.308.308","article-title":"Molecular genetic analysis of porcine von Willebrand disease: tight linkage to the von Willebrand factor locus.","volume":"72","author":"Bahou","year":"1988","journal-title":"Blood"},{"key":"2021042013282762400_bib4","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/S1079-9796(03)00033-0","article-title":"Molecular defects in type 3 von Willebrand disease: updated results from 40 multiethnic patients.","volume":"30","author":"Baronciani","year":"2003","journal-title":"Blood Cells Mol. Dis."},{"key":"2021042013282762400_bib5","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.2460\/ajvr.1973.34.11.1405","article-title":"Tests of hemostasis in swine: normal values and values in pigs affected with von Willebrand\u2019s disease.","volume":"34","author":"Bowie","year":"1973","journal-title":"Am. J. Vet. Res."},{"key":"2021042013282762400_bib6","doi-asserted-by":"crossref","first-page":"3055","DOI":"10.1161\/01.ATV.19.12.3055","article-title":"In vivo regulation of von Willebrand factor synthesis: von Willebrand factor production in endothelial cells after lung transplantation between normal pigs and von Willebrand factor-deficient pigs.","volume":"19","author":"Brouland","year":"1999","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2021042013282762400_bib7","doi-asserted-by":"crossref","first-page":"667","DOI":"10.3324\/haematol.2012.077263","article-title":"Principles of care for the diagnosis and treatment of von Willebrand disease.","volume":"98","author":"Castaman","year":"2013","journal-title":"Haematologica"},{"key":"2021042013282762400_bib8","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1006\/mgme.1999.2864","article-title":"Alu repeats and human disease.","volume":"67","author":"Deininger","year":"1999","journal-title":"Mol. Genet. Metab."},{"key":"2021042013282762400_bib9","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1182\/blood.V53.4.712.712","article-title":"Inheritance of porcine von Willebrand\u2019s disease: study of a kindred of over 700 pigs.","volume":"53","author":"Fass","year":"1979","journal-title":"Blood"},{"key":"2021042013282762400_bib10","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1182\/asheducation.V2014.1.531.3883265","article-title":"New insights into genotype and phenotype of VWD.","volume":"1","author":"Flood","year":"2014","journal-title":"Hematology Am. Soc. Hematol. Educ. Program"},{"key":"2021042013282762400_bib11","doi-asserted-by":"crossref","first-page":"437","DOI":"10.2460\/javma.1987.190.04.437","article-title":"A bleeding disorder (von Willebrand\u2019s disease) in a Himalayan cat.","volume":"190","author":"French","year":"1987","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"2021042013282762400_bib12","doi-asserted-by":"crossref","first-page":"99","DOI":"10.5315\/wjh.v2.i4.99","article-title":"Pseudohemophilia of Erik von Willebrand caused by homozygous one nucleotide deletion in exon 18 of the VW-factor gene.","volume":"2","author":"Gadisseur","year":"2013","journal-title":"World J. Hepatol."},{"key":"2021042013282762400_bib13"},{"key":"2021042013282762400_bib14","doi-asserted-by":"crossref","first-page":"2087","DOI":"10.1073\/pnas.71.5.2087","article-title":"Von Willebrand factor: gene dosage relationships and transfusion response in bleeder swine: a new bioassay.","volume":"71","author":"Griggs","year":"1974","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021042013282762400_bib15","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1016\/j.placenta.2015.06.001","article-title":"Changes in endometrial ezrin and cytokeratin 18 expression during bovine implantation and in caruncular endometrial spheroids in vitro.","volume":"36","author":"Haeger","year":"2015","journal-title":"Placenta"},{"key":"2021042013282762400_bib16","doi-asserted-by":"crossref","first-page":"217","DOI":"10.3181\/00379727-48-13275P","article-title":"A hemophilia-like disease in swine.","volume":"48","author":"Hogan","year":"1941","journal-title":"Proc. Soc. Exp. Biol. Med."},{"key":"2021042013282762400_bib17","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/humu.1380010103","article-title":"Partial gene duplication as a cause of human disease.","volume":"1","author":"Hu","year":"1992","journal-title":"Hum. Mutat."},{"key":"2021042013282762400_bib18","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1093\/nar\/gkw010","article-title":"Mechanism and regulation of the nonsense-mediated decay pathway.","volume":"44","author":"Hug","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"2021042013282762400_bib19","doi-asserted-by":"crossref","first-page":"e92575","DOI":"10.1371\/journal.pone.0092575","article-title":"Genetic heterogeneity in a large cohort of Indian type 3 von Willebrand disease patients.","volume":"9","author":"Kasatkar","year":"2014","journal-title":"PLoS One"},{"key":"2021042013282762400_bib20","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1093\/bioinformatics\/btm091","article-title":"Enhancements and modifications of primer design program Primer3.","volume":"23","author":"Koressaar","year":"2007","journal-title":"Bioinformatics"},{"key":"2021042013282762400_bib21","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1093\/bioinformatics\/btp324","article-title":"Fast and accurate short read alignment with Burrows-Wheeler transform.","volume":"25","author":"Li","year":"2009","journal-title":"Bioinformatics"},{"key":"2021042013282762400_bib22","first-page":"3","article-title":"The molecular genetics of von Willebrand disease.","volume":"1","author":"Lillicrap","year":"2005","journal-title":"Haematol. Rep."},{"key":"2021042013282762400_bib23","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(\u2212Delta Delta C(T)) Method.","volume":"25","author":"Livak","year":"2001","journal-title":"Methods."},{"key":"2021042013282762400_bib24","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1111\/j.1365-2516.2009.02025.x","article-title":"Bleeding symptoms and laboratory correlation in patients with severe von Willebrand disease.","volume":"15","author":"Metjian","year":"2009","journal-title":"Haemophilia"},{"key":"2021042013282762400_bib25","doi-asserted-by":"crossref","first-page":"2029","DOI":"10.1182\/blood-2014-08-528398","article-title":"Diagnostic approach to von Willebrand disease.","volume":"125","author":"Ng","year":"2015","journal-title":"Blood"},{"key":"2021042013282762400_bib26","first-page":"461238","article-title":"Porcine and canine von Willebrand factor and von Willebrand disease: hemostasis, thrombosis, and atherosclerosis studies.","volume-title":"Thrombosis","author":"Nichols","year":"2010"},{"key":"2021042013282762400_bib27","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1089\/cmb.2013.0084","article-title":"Assembling single-cell genomes and mini-metagenomes from chimeric MDA products.","volume":"20","author":"Nurk","year":"2013","journal-title":"J. Comput. Biol."},{"key":"2021042013282762400_bib28","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1055\/s-0037-1615197","article-title":"Identification of mutations in the canine von Willebrand factor gene associated with type III von-Willebrand-disease.","volume":"80","author":"Rieger","year":"1998","journal-title":"Thromb. Haemost."},{"key":"2021042013282762400_bib29","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.1038\/ng.185","article-title":"A cis-acting regulatory mutation causes premature hair graying and susceptibility to melanoma in the horse.","volume":"40","author":"Rosengren Pielberg","year":"2008","journal-title":"Nat. Genet."},{"key":"2021042013282762400_bib30","first-page":"585","article-title":"An experimental model for testing von Willebrand factor function: successful SLA-matched crossed bone marrow transplantations between normal and von Willebrand pigs.","volume":"24","author":"Roussi","year":"1996","journal-title":"Exp. Hematol."},{"key":"2021042013282762400_bib31","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1097\/00001721-199806000-00009","article-title":"Effects of human recombinant, plasma-derived and porcine von Willebrand factor in pigs with severe von Willebrand disease.","volume":"9","author":"Roussi","year":"1998","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"2021042013282762400_bib32","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1146\/annurev.biochem.67.1.395","article-title":"Biochemistry and genetics of von Willebrand factor.","volume":"67","author":"Sadler","year":"1998","journal-title":"Annu. Rev. Biochem."},{"key":"2021042013282762400_bib33","doi-asserted-by":"crossref","first-page":"2103","DOI":"10.1111\/j.1538-7836.2006.02146.x","article-title":"Working Party on von Willebrand Disease Classification. Update on the pathophysiology and classification of von Willebrand disease: a report of the subcommittee on von Willebrand Factor.","volume":"4","author":"Sadler","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"2021042013282762400_bib34","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1038\/ng.2007.4","article-title":"Duplication of FGF3, FGF4, FGF19 and ORAOV1 causes hair ridge and predisposition to dermoid sinus in Ridgeback dogs.","volume":"39","author":"Salmon Hillbertz","year":"2007","journal-title":"Nat. Genet."},{"key":"2021042013282762400_bib35","doi-asserted-by":"crossref","first-page":"2390","DOI":"10.1073\/pnas.95.5.2390","article-title":"The partial tandem duplication of ALL1 (MLL) is consistently generated by Alu-mediated homologous recombination in acute myeloid leukemia.","volume":"95","author":"Strout","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2021042013282762400_bib36","doi-asserted-by":"crossref","first-page":"1637","DOI":"10.1182\/blood.V94.5.1637","article-title":"Evidence for extracellular processing of pro-von Willebrand factor after infusion in animals with and without severe von Willebrand disease.","volume":"94","author":"Turecek","year":"1999","journal-title":"Blood"},{"key":"2021042013282762400_bib37"},{"key":"2021042013282762400_bib38","first-page":"10","article-title":"Mutation causing von Willebrand\u2019s disease in Scottish Terriers.","volume":"14","author":"Venta","year":"2000","journal-title":"J. Vet. Intern. Med."},{"key":"2021042013282762400_bib39","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1093\/hmg\/1.9.767","article-title":"A single cytosine deletion in exon 18 of the von Willebrand factor gene is the most common mutation in Swedish vWD type III patients.","volume":"1","author":"Zhang","year":"1992","journal-title":"Hum. Mol. Genet."}],"container-title":["G3 Genes|Genomes|Genetics"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/g3journal\/article-pdf\/8\/2\/577\/37185470\/g3journal0577.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/g3journal\/article-pdf\/8\/2\/577\/37185470\/g3journal0577.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,28]],"date-time":"2025-06-28T04:05:58Z","timestamp":1751083558000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/g3journal\/article\/8\/2\/577\/6028121"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,2,1]]},"references-count":39,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2018,2,1]]}},"URL":"https:\/\/doi.org\/10.1534\/g3.117.300432","relation":{},"ISSN":["2160-1836"],"issn-type":[{"type":"electronic","value":"2160-1836"}],"subject":[],"published-other":{"date-parts":[[2018,2,1]]},"published":{"date-parts":[[2018,2,1]]}}}