{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:24:30Z","timestamp":1768508670640,"version":"3.49.0"},"posted":{"date-parts":[[2023,11,29]]},"group-title":"Molecular Biology","reference-count":44,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,11,29]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  <jats:italic>Ixodes scapularis<\/jats:italic>\n                  is an important vector of many pathogens, including the causative agent of Lyme disease, tick-borne encephalitis, and anaplasmosis. The study of gene function in\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  and other ticks has been hampered by the lack of genetic tools, such as an inducible promoter to permit temporal control over transgenes encoding protein or double-stranded RNA expression. Studies of vector-pathogen relationships would also benefit from the capability to activate anti-pathogen genes at different times during pathogen infection and dissemination. We have characterized an intergenic sequence upstream of the heat shock protein 70 (HSP70) gene that can drive\n                  <jats:italic>Renilla<\/jats:italic>\n                  luciferase expression and mCherry fluorescence in the\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  cell line ISE6. In another construct, we replaced the\n                  <jats:italic>Drosophila melanogaster<\/jats:italic>\n                  minimal HSP70 promoter in the synthetic 3xP3 promoter with a minimal portion of the\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  HSP70 promoter and generated an\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  specific 3xP3 (Is3xP3) promoter. Both promoter constructs, IsHSP70 and Is3xP3, allow for heat-inducible expression of mCherry fluorescence in ISE6 cells with an approximately 10-fold increase in the percentage of fluorescent positive cells upon exposure to a 2 h heat shock. These promoters described here will be valuable tools for gene function studies and temporal control of gene expression, including anti-pathogen genes.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"569248v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2023.11.29.569248","type":"posted-content","created":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T22:50:10Z","timestamp":1701298210000},"source":"Crossref","is-referenced-by-count":0,"title":["Validation of a heat-inducible\n                  <i>Ixodes scapularis<\/i>\n                  HSP70 promoter and developing a tick-specific 3xP3 promoter sequence in ISE6 cells"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5698-9335","authenticated-orcid":false,"given":"Michael","family":"Pham","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hans-Heinrich","family":"Hoffmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7935-2249","authenticated-orcid":false,"given":"Timothy J.","family":"Kurtti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Randeep","family":"Chana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Omar","family":"Garcia-Cruz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simindokht","family":"Aliabadi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7095-1037","authenticated-orcid":false,"given":"Monika","family":"Gulia-Nuss","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023120400200546000_2023.11.29.569248v1.1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2583.2010.01005.x"},{"key":"2023120400200546000_2023.11.29.569248v1.2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ento.46.1.317"},{"key":"2023120400200546000_2023.11.29.569248v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/46463"},{"key":"2023120400200546000_2023.11.29.569248v1.4","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2583.2011.01116.x"},{"key":"2023120400200546000_2023.11.29.569248v1.5","unstructured":"Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases (NCEZID), Division of Vector-Borne Diseases. 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