{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T03:59:07Z","timestamp":1782273547120,"version":"3.54.5"},"reference-count":52,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2021,8,18]],"date-time":"2021-08-18T00:00:00Z","timestamp":1629244800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 NS086134"],"award-info":[{"award-number":["R01 NS086134"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 NS064330"],"award-info":[{"award-number":["R01 NS064330"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2021,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Co\u2010expression of multiple genes in single vectors has achieved varying degrees of success by employing two promoters and\/or application of viral 2A\u2010peptide or the internal ribosome entry\u2010site (IRES). However, promoter interference, potential functional\u2010interruption of expressed\u2010proteins by 2A\u2010generated residual peptides or weaker translation of IRES\u2010mediated downstream genes has curtailed their utilization. Thus, there is the need for single vectors that robustly express multiple proteins for enhanced gene therapy applications.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We engineered lentiviral\u2010vectors for dual\u2010cassette expression of green fluorescent protein and mCherry in uni\u2010 or bidirectional architectures using the short\u2010version (Es) of elongation factor 1\u03b1 (EF) promoter and simian virus 40 promoter (Sv). The regulatory function of a core fragment (cC) from human cytomegalovirus promoter was investigated with cell\u2010lineage specificity in NIH3T3 (fibroblast) and hematopoietic cell lines U937 (monocyte\/macrophage), LCL (lymphoid), DAMI (megakaryocyte) and MEL (erythroid).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The cC element in reverse\u2010orientation not only boosted upstream Es promoter to levels comparable to full\u2010length EF in DAMI, U937 and 3T3 cells, but also blocked the suppression of downstream Sv promoter by Es in U937 and 3T3 cells with further improved Sv activity in DAMI cells. Such lineage\u2010restricted up\u2010regulation is likely attributed to two protein\u2010binding domains of cC and diverse expression of related factors in different cell types for enhancer and terminator activities, but not spacing function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Such a newly developed dual\u2010cassette vector could be advantageous, particularly in hematopoietic cell\u2010mediated gene\/cancer therapy, by allowing for independent and robust co\u2010expression of therapeutic gene(s) and\/or a selectable gene or imaging marker in the same cells.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.3380","type":"journal-article","created":{"date-parts":[[2021,7,28]],"date-time":"2021-07-28T02:54:05Z","timestamp":1627440845000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["New function of a well\u2010known promoter: Enhancer activity of minimal CMV promoter enables efficient dual\u2010cassette transgene expression"],"prefix":"10.1002","volume":"23","author":[{"given":"Michael K. 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