Abstract
This protocol describes the design, cloning, and in vitro screening of artificial microRNAs (miRNAs) to silence alpha-1 antitrypsin (AAT). This method would be of interest to silence AAT in a variety of in vitro or in vivo models, and prevalidated sequences against human AAT are provided. This simple 5-day protocol may more generally be used to design artificial miRNAs against any transcript.
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References
Mueller C, Tang Q, Gruntman A, Blomenkamp K, Teckman J, Song L, Zamore PD, Flotte TR (2012) Sustained miRNA-mediated knockdown of mutant AAT with simultaneous augmentation of wild-type AAT has minimal effect on global liver miRNA profiles. Mol Ther 20(3):590–600. doi:10.1038/mt.2011.292. mt2011292 [pii]
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Borel, F., Mueller, C. (2017). Design, Cloning, and In Vitro Screening of Artificial miRNAs to Silence Alpha-1 Antitrypsin. In: Borel, F., Mueller, C. (eds) Alpha-1 Antitrypsin Deficiency . Methods in Molecular Biology, vol 1639. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7163-3_10
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DOI: https://doi.org/10.1007/978-1-4939-7163-3_10
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7161-9
Online ISBN: 978-1-4939-7163-3
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