{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T08:57:45Z","timestamp":1762505865640},"reference-count":39,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2012,1,18]],"date-time":"2012-01-18T00:00:00Z","timestamp":1326844800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Oral Diseases"],"published-print":{"date-parts":[[2012,7]]},"abstract":"<jats:p><jats:italic>Oral Diseases<\/jats:italic> (2012) <jats:bold>18<\/jats:bold>, 477\u2013484<\/jats:p><jats:p><jats:bold>Objective:\u2002<\/jats:bold> ELAV\u2010like proteins regulate mRNA stability and\/or translation. We evaluated whether inclusion of binding sites for ELAV\u2010like HuR proteins in vector cassettes could improve transgene expression in the salivary gland.<\/jats:p><jats:p><jats:bold>Methods:\u2002<\/jats:bold> Western blots and immunofluorescence staining were used to determine whether HuR protein was expressed in salivary cells and tissue. HuR binding sites were inserted into the pACEF1\u03b1\u2013luc\u2010BGH expression plasmid. Cell lines were transfected with plasmids <jats:italic>in vitro<\/jats:italic> and luciferase expression measured. Rat submandibular glands were transfected <jats:italic>in vivo<\/jats:italic> with plasmids containing ELAV\u2010like HuR protein\u2010binding sites. An adenoviral vector with p53 ELAV\u2010like HuR protein\u2010binding site was generated and also tested <jats:italic>in vivo<\/jats:italic>. Four unique 29mer HuR shRNA constructs were used in A5 cells to evaluate whether there was a specific interaction between HuR protein and the p53 HuR protein\u2010binding site.<\/jats:p><jats:p><jats:bold>Results:\u2002<\/jats:bold> Salivary cells express HuR protein. Inclusion of the p53 ELAV\u2010like HuR protein\u2010binding site resulted in high luciferase activity in salivary cells <jats:italic>in vitro<\/jats:italic>, with similar results <jats:italic>in vivo<\/jats:italic>. <jats:italic>In vitro<\/jats:italic> shRNA data demonstrated that the high luciferase activity was mediated by the interaction between HuR protein and the p53 HuR protein\u2010binding site. The AdEF1\u03b1\u2010luc\u2010p53BGH, including this binding site, mediated very high luciferase activity, \u223c4\u2010fold that seen with the CMV promoter, in rat submandibular glands.<\/jats:p><jats:p><jats:bold>Conclusions:\u2002<\/jats:bold> Including the p53 ELAV\u2010like protein\u2010binding site in transgene cassettes may enhance therapeutic vectors intended for use with salivary glands.<\/jats:p>","DOI":"10.1111\/j.1601-0825.2011.01895.x","type":"journal-article","created":{"date-parts":[[2011,12,22]],"date-time":"2011-12-22T01:04:05Z","timestamp":1324515845000},"page":"477-484","source":"Crossref","is-referenced-by-count":6,"title":["Including the p53 ELAV\u2010like protein\u2010binding site in vector cassettes enhances transgene expression in rat submandibular 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