{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T16:01:24Z","timestamp":1790438484587,"version":"4.1.0"},"reference-count":4,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Single\u2010nucleotide polymorphisms (SNPs) may influence gene expression and contribute to interindividual variability in immune responses and disease susceptibility. Interleukin\u201033 (IL33), a cytokine involved in inflammation, immune regulation, epithelial barrier homeostasis, and tissue remodeling, has been implicated in allergic, autoimmune, inflammatory, and neoplastic disorders. The\n                    <jats:italic>IL33<\/jats:italic>\n                    rs1929992 (T\u00a0&gt;\u00a0C) polymorphism, located in intron 3, has been associated with altered IL33 expression, with the T allele linked to increased cytokine production compared with the C allele. This article describes a standardized sequence\u2010specific primer polymerase chain reaction (PCR\u2010SSP) protocol for genotyping the human\n                    <jats:italic>IL33<\/jats:italic>\n                    rs1929992 polymorphism, with validation by Sanger sequencing. The protocol includes optimization of annealing temperature, MgCl\n                    <jats:sub>2<\/jats:sub>\n                    concentration, primer balance, deoxynucleotide triphosphates (dNTPs), and DNA input to ensure amplification specificity and reproducibility. Allele\u2010specific amplification is achieved using forward primers specific for the T and C alleles in independent reactions and analyzed by agarose gel electrophoresis, with the\n                    <jats:italic>human growth hormone<\/jats:italic>\n                    (\n                    <jats:italic>HGH<\/jats:italic>\n                    ) gene used as an internal control. This protocol provides a simple, reproducible, and accessible method for rs1929992 genotyping, suitable for population\u2010based and genetic association studies involving immune\u2010mediated and inflammatory diseases. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : PCR\u2010SSP genotyping of\n                    <jats:italic>IL33<\/jats:italic>\n                    rs1929992\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Extraction of genomic DNA by salting\u2010out\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70455","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T16:46:31Z","timestamp":1788194791000},"source":"Crossref","is-referenced-by-count":0,"title":["Sequence\u2010Specific Primer Polymerase Chain Reaction Genotyping of the Human\n                    <i>IL33<\/i>\n                    Polymorphism rs1929992 (T\u00a0&gt;\u00a0C) with Sanger Sequencing Validation"],"prefix":"10.1002","volume":"6","author":[{"given":"Al\u00e9ia Harumi Uchibaba","family":"Yamanaka","sequence":"first","affiliation":[{"name":"Department of Basic Health Sciences (DBS), Laboratory of Immunogenetics (Imunogen\u2010UEM) State University of Maring\u00e1 Maring\u00e1 Paran\u00e1 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guilherme","family":"Castellani","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM\u201056), Hospital das Cl\u00ednicas, Faculdade de Medicina da Universidade de S\u00e3o Paulo (HCFMUSP) University of S\u00e3o Paulo S\u00e3o Paulo S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matheus","family":"Braga","sequence":"additional","affiliation":[{"name":"Department of Basic Health Sciences (DBS), Laboratory of Immunogenetics (Imunogen\u2010UEM) State University of Maring\u00e1 Maring\u00e1 Paran\u00e1 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josiane Bazzo","family":"de Alencar","sequence":"additional","affiliation":[{"name":"Department of Basic Health Sciences (DBS), Laboratory of Immunogenetics (Imunogen\u2010UEM) State University of Maring\u00e1 Maring\u00e1 Paran\u00e1 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Let\u00edcia Cristina de Almeida","family":"Silva","sequence":"additional","affiliation":[{"name":"Department of Basic Health Sciences (DBS), Laboratory of Immunogenetics (Imunogen\u2010UEM) State University of Maring\u00e1 Maring\u00e1 Paran\u00e1 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quirino Alves","family":"de Lima Neto","sequence":"additional","affiliation":[{"name":"Department of Basic Health Sciences (DBS), Laboratory of Immunogenetics (Imunogen\u2010UEM) State University of Maring\u00e1 Maring\u00e1 Paran\u00e1 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpz1.70390"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471140864.psa04ks52"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/19.2.408"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70455","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70455","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70455","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T15:55:16Z","timestamp":1790438116000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70455"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,31]]},"references-count":4,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/cpz1.70455"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70455","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,31]]},"article-number":"e70455"}}