Abstract
Alternative polyadenylation is increasingly being recognized as an important layer of gene regulation. Antisense-mediated modulation of alternative polyadenylation represents an attractive strategy for the regulation of gene expression as well as potential therapeutic applications. In this chapter, we describe methods to upregulate the functional Kv11.1 isoform expression by blocking intronic polyadenylation signal sequences with antisense morpholinos.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Shi Y (2012) Alternative polyadenylation: new insights from global analyses. RNA 18:2105–2117
Tian B, Manley JL (2013) Alternative cleavage and polyadenylation: the long and short of it. Trends Biochem Sci 38:312–320
Elkon R, Ugalde AP, Agami R (2013) Alternative cleavage and polyadenylation: extent, regulation and function. Nat Rev Genet 14:496–506
Hollerer I, Grund K, Hentze MW, Kulozik AE (2014) mRNA 3′end processing: A tale of the tail reaches the clinic. EMBO Mol Med 6:16–26
Gong Q, Stump MR, Deng V, Zhang L, Zhou Z (2014) Identification of Kv11.1 isoform switch as a novel pathogenic mechanism of long QT syndrome. Circ Cardiovasc Genet 76:26–32
Bennett CF, Swayze EE (2010) RNA targeting therapeutics: molecular mechanisms of antisense oligonucleotides as a therapeutic platform. Annu Rev Pharmacol Toxicol 50:259–293
Vickers TA, Wyatt JR, Burckin T, Bennett CF, Freier SM (2001) Fully modified 2′ MOE oligonucleotides redirect polyadenylation. Nucleic Acids Res 29:1293–1299
Vorlová S, Rocco G, Lefave CV, Jodelka FM, Hess K, Hastings ML, Henke E, Cartegni L (2011) Induction of antagonistic soluble decoy receptor tyrosine kinases by intronic polyA activation. Mol Cell 43:927–939
Gong Q, Stump MR, Zhou Z (2014) Upregulation of functional Kv11.1 isoform expression by inhibition of intronic polyadenylation with antisense morpholino oligonucleotides. J Mol Cell Cardiol 76:26–32
Raz V, Buijze H, Raz Y, Verwey N, Anvar SY, Aartsma-Rus A, van der Maarel SM (2014) A novel feed-forward loop between ARIH2 E3-ligase and PABPN1 regulates aging-associated muscle degeneration. Am J Pathol 184:1119–1131
Warmke JW, Ganetzky B (1994) A family of potassium channel genes related to eag in Drosophila and mammals. Proc Natl Acad Sci USA 91:3438–3442
Sanguinetti MC, Jiang C, Curran ME, Keating MT (1995) A mechanistic link between an inherited and an acquired cardiac arrhythmia: HERG encodes the IKr potassium channel. Cell 81:299–307
Trudeau MC, Warmke JW, Ganetzky B, Robertson GA (1995) HERG, a human inward rectifier in the voltage-gated potassium channel family. Science 269:92–95
Zhou Z, Gong Q, Ye B, Fan Z, Makielski JC, Robertson GA, January CT (1998) Properties of HERG channels stably expressed in HEK 293 cells studied at physiological temperature. Biophys J 74:230–241
Curran ME, Splawski I, Timothy KW, Vincent GM, Green ED, Keating MT (1995) A molecular basis for cardiac arrhythmia: HERG mutations cause long QT syndrome. Cell 80:795–803
Kannankeril P, Roden DM, Darbar D (2010) Drug-induced long QT syndrome. Pharmacol Rev 62:760–781
Gong Q, Stump MR, Dunn AR, Deng V, Zhou Z (2010) Alternative splicing and polyadenylation contribute to the generation of hERG1 C-terminal isoforms. J Biol Chem 285:32233–32241
Kupershmidt S, Snyders DJ, Raes A, Roden DM (1998) A K+ channel splice variant common in human heart lacks a C-terminal domain required for expression of rapidly activating delayed rectifier current. J Biol Chem 273:27231–27235
Acknowledgments
This study was supported in part by NIH grant HL68854.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Gong, Q., Zhou, Z. (2017). Regulation of Isoform Expression by Blocking Polyadenylation Signal Sequences with Morpholinos. In: Moulton, H., Moulton, J. (eds) Morpholino Oligomers. Methods in Molecular Biology, vol 1565. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6817-6_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6817-6_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6815-2
Online ISBN: 978-1-4939-6817-6
eBook Packages: Springer Protocols