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- W4200425114 abstract "Manipulating expression of large genes (>6 kb) in adult cardiomyocytes is challenging because these cells are only efficiently transduced by viral vectors with a 4-7 kb packaging capacity. This limitation impedes understanding structure-function mechanisms of important proteins in heart. L-type calcium channels (LTCCs) regulate diverse facets of cardiac physiology including excitation-contraction coupling, excitability, and gene expression. Many important questions about how LTCCs mediate such multidimensional signaling are best resolved by manipulating expression of the 6.6 kb pore-forming α1C-subunit in adult cardiomyocytes. Here, we use split-intein-mediated protein transsplicing to reconstitute LTCC α1C-subunit from two distinct halves, overcoming the difficulty of expressing full-length α1C in cardiomyocytes. Split-intein-tagged α1C fragments encoding dihydropyridine-resistant channels were incorporated into adenovirus and reconstituted in cardiomyocytes. Similar to endogenous LTCCs, recombinant channels targeted to dyads, triggered Ca(2+) transients, associated with caveolin-3, and supported β-adrenergic regulation of excitation-contraction coupling. This approach lowers a longstanding technical hurdle to manipulating large proteins in cardiomyocytes. PMID: 24003157 Funding information This work was supported by: NHLBI NIH HHS, United States Grant ID: R01 HL 069911 NHLBI NIH HHS, United States Grant ID: R01 HL069911" @default.
- W4200425114 created "2021-12-31" @default.
- W4200425114 creator A5021749666 @default.
- W4200425114 date "2013-09-27" @default.
- W4200425114 modified "2023-09-28" @default.
- W4200425114 title "Faculty Opinions recommendation of Manipulating L-type calcium channels in cardiomyocytes using split-intein protein transsplicing." @default.
- W4200425114 doi "https://doi.org/10.3410/f.718098602.793483607" @default.
- W4200425114 hasPublicationYear "2013" @default.
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