Abstract
This chapter describes the potential use of viral-mediated gene transfer in the central nervous system for the silencing of gene expression using RNA interference in the context of Huntington’s disease (HD). Protocols provided here describe the design of small interfering RNAs, their encoding in lentiviral vectors (LVs) and viral production, as well as procedures for their stereotaxic injection in the rodent brain.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wang YL, Liu W, Wada E et al (2005) Clinico-pathological rescue of a model mouse of Huntington’s disease by siRNA. Neurosci Res 53:241–249
DiFiglia M, Sena-Esteves M, Chase K et al (2007) Therapeutic silencing of mutant huntingtin with siRNA attenuates striatal and cortical neuropathology and behavioral deficits. Proc Natl Acad Sci USA 104:17204–17209
Samuel-Abraham S, Leonard JN (2010) Staying on message: design principles for controlling nonspecific responses to siRNA. FEBS J 277:4828–4836
Abbas-Terki T, Blanco-Bose W, Deglon N et al (2002) Lentiviral-mediated RNA interference. Hum Gene Ther 13:2197–2201
Deglon N, Tseng JL, Bensadoun JC et al (2000) Self-inactivating lentiviral vectors with enhanced transgene expression as potential gene transfer system in Parkinson’s disease. Hum Gene Ther 11:179–190
Naldini L, Blomer U, Gallay P et al (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272:263–267
Abordo-Adesida E, Follenzi A, Barcia C et al (2005) Stability of lentiviral vector-mediated transgene expression in the brain in the presence of systemic antivector immune responses. Hum Gene Ther 16:741–751
Grimm D, Streetz KL, Jopling CL et al (2006) Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways. Nature 441:537–541
Drouet V, Perrin V, Hassig R et al (2009) Sustained effects of nonallele-specific Huntingtin silencing. Ann Neurol 65:276–285
Boudreau RL, McBride JL, Martins I et al (2009) Nonallele-specific silencing of mutant and wild-type huntingtin demonstrates therapeutic efficacy in Huntington’s disease mice. Mol Ther 17:1053–1063
Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553
Grimm D (2009) Small silencing RNAs: state-of-the-art. Adv Drug Deliv Rev 61:672–703
Snove O Jr, Rossi JJ (2006) Expressing short hairpin RNAs in vivo. Nat Methods 3:689–695
Walton SP, Wu M, Gredell JA et al (2010) Designing highly active siRNAs for therapeutic applications. FEBS J 277:4806–4813
Liu YP, Vink MA, Westerink JT et al (2010) Titers of lentiviral vectors encoding shRNAs and miRNAs are reduced by different mechanisms that require distinct repair strategies. RNA 16:1328–1339
Follenzi A, Ailles LE, Bakovic S et al (2000) Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nat Genet 25:217–222
Zufferey R, Donello JE, Trono D et al (1999) Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J Virol 73:2886–2892
Paxinos G, Watson C (2004) The rat brain in stereotaxic coordinates (New York, Academic press)
Hottinger AF, Azzouz M, Deglon N et al (2000) Complete and long-term rescue of lesioned adult motoneurons by lentiviral-mediated expression of glial cell line-derived neurotrophic factor in the facial nucleus. J Neurosci 20:5587–5593
Barde I, Salmon P, Trono D (2010) Production and titration of lentiviral vectors. Curr Protoc Neurosci Chapter 4, Unit 4 21
Kutner RH, Zhang XY, Reiser J (2009) Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors. Nat Protoc 4:495–505
Franklin B, Paxinos G (1996) The mouse brain in stereotaxic coordinates (New York)
Farson D, Witt R, McGuinness R et al (2001) A new-generation stable inducible packaging cell line for lentiviral vectors. Hum Gene Ther 12:981–997
Yang S, Delgado R, King SR et al (1999) Generation of retroviral vector for clinical studies using transient transfection. Hum Gene Ther 10:123–132
Boudreau RL, Monteys AM, Davidson BL (2008) Minimizing variables among hairpin-based RNAi vectors reveals the potency of shRNAs. RNA 14:1834–1844
Khan AA, Betel D, Miller ML et al (2009) Transfection of small RNAs globally perturbs gene regulation by endogenous microRNAs. Nat Biotechnol 27:549–555
Grimm D, Wang L, Lee JS et al (2010) Argonaute proteins are key determinants of RNAi efficacy, toxicity, and persistence in the adult mouse liver. J Clin Invest 120:3106–3119
Chen G, Kronenberger P, Teugels E et al (2011) Influence of RT-qPCR primer position on EGFR interference efficacy in lung cancer cells. Biol Proced Online 13:1
Shepard AR, Jacobson N, Clark AF (2005) Importance of quantitative PCR primer location for short interfering RNA efficacy determination. Anal Biochem 344:287–288
Hahn P, Schmidt C, Weber M et al (2004) RNA interference: PCR strategies for the quantification of stable degradation-fragments derived from siRNA-targeted mRNAs. Biomol Eng 21:113–117
Boudreau RL, Martins I, Davidson BL (2009) Artificial microRNAs as siRNA shuttles: improved safety as compared to shRNAs in vitro and in vivo. Mol Ther 17:169–175
Regulier E, Zala D, Aebischer P et al (2004) Lentiviral-mediated gene transfer to model triplet repeat disorders. Methods Mol Biol 277:199–213
Rodrigues T, Carrondo MJ, Alves PM et al (2007) Purification of retroviral vectors for clinical application: biological implications and technological challenges. J Biotechnol 127:520–541
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer New York
About this protocol
Cite this protocol
Cambon, K., Déglon, N. (2013). Lentiviral-Mediated Gene Transfer of siRNAs for the Treatment of Huntington’s Disease. In: Kohwi, Y., McMurray, C. (eds) Trinucleotide Repeat Protocols. Methods in Molecular Biology, vol 1010. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-411-1_7
Download citation
DOI: https://doi.org/10.1007/978-1-62703-411-1_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-410-4
Online ISBN: 978-1-62703-411-1
eBook Packages: Springer Protocols