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Viral Vector Approaches in Neurobiology and Brain Diseases pp 11–25Cite as

Recombinant Adenovirus in Neurobiology

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Part of the book series: Neuromethods ((NM,volume 82))

Abstract

Recombinant adenoviruses have been extensively applied in basic research and gene therapy. Recently, there is an increase in neurobiological studies using recombinant adenovirus technology to manipulate gene expression in neurological systems. Recombinant adenovirus technology provides a useful tool for investigating the function of a gene of interest, neurocircuitries and the delivery of genes for therapy. In order to take full advantage of this technique, it is important to understand the strategies for the design and application of the virus. The goal of this chapter is to provide a practical protocol on the application of recombinant adenovirus for neurobiological studies. A simplified recombinant adenoviral system, AdEasy, will be applied to generate recombinant adenoviruses for the expression and knockdown of the gene of interest. We discuss the construction of vectors for gene expression and siRNAs, the generation of adenoviral recombinants, and the production of high-titer adenovirus. Furthermore, a procedure for injection of the adenovirus using a stereotaxic apparatus is discussed.

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References

  1. Choi JW, Lee JS, Kim SW, Yun CO (2012) Evolution of oncolytic adenovirus for cancer treatment. Adv Drug Deliv Rev 64:720–729

    Article  PubMed  CAS  Google Scholar 

  2. Gomez-Manzano C, Jiang H, Alonso M, Yung WK, Fueyo J (2012) Gene therapy. In: Vinken PJ, Bruyn GW (eds) Handbook of clinical neurology, vol 104., pp 331–338

    Google Scholar 

  3. Kamimura K, Suda T, Zhang G, Liu D (2011) Advances in gene delivery systems. Pharmaceut Med 25:293–306

    PubMed  Google Scholar 

  4. Yao XL, Nakagawa S, Gao JQ (2011) Current targeting strategies for adenovirus vectors in cancer gene therapy. Curr Cancer Drug Targets 11:810–825

    Article  PubMed  CAS  Google Scholar 

  5. Li Q, Luo T, Jiang X, Wang J (2012) Anxiolytic effects of 5-HTA receptors and anxiogenic effects of 5-HTC receptors in the amygdala of mice. Neuropharmacology 62:474–484

    Article  PubMed  CAS  Google Scholar 

  6. Li Q, Holmes A, Ma L, Van de Kar LD, Garcia F, Murphy DL (2004) Medial hypothalamic 5-hydroxytryptamine (5-HT)1A receptors regulate neuroendocrine responses to stress and exploratory locomotor activity: application of recombinant adenovirus containing 5-HT1A sequences. J Neurosci 24:10868–10877

    Article  PubMed  CAS  Google Scholar 

  7. Rossi DV, Dai Y, Thomas P, Carrasco GA, DonCarlos LL, Muma NA, Li Q (2010) Estradiol-induced desensitization of 5-HT(1A) receptor signaling in the paraventricular nucleus of the hypothalamus is independent of estrogen receptor-beta. Psychoneuroendocrinology 35:1023–1033

    Article  PubMed  CAS  Google Scholar 

  8. McAllister CE, Creech RD, Kimball PA, Muma NA, Li Q (2012) GPR30 is necessary for Q1 estradiol-induced desensitization of 5-HT1A receptor signaling in the paraventricular nucleus of the rat hypothalamus. Psychoneuroendocrinology 37:1248–1260

    Article  PubMed  CAS  Google Scholar 

  9. Cazzin C, Zanderigo F, Costantini VJ, Zambello E, Ring CJ, Morrison AD, Caberlotto L, Kew JN (2010) Adenoviral-mediated Cre expression effectively suppresses GlyT1 binding in the thalamic area of GlyT1 conditional knock-out mice. J Neurosci Methods 193:254–263

    Article  PubMed  CAS  Google Scholar 

  10. Wang Y, Krushel LA, Edelman GM (1996) Targeted DNA recombination in vivo using an adenovirus carrying the cre recombinase gene. Proc Natl Acad Sci USA 93:3932–3936

    Article  PubMed  CAS  Google Scholar 

  11. Kim ML, Han S, Lee SB, Kim JH, Ahn HK, Huh Y (2010) Evaluation of recombinant adenovirus-mediated gene delivery for expression of tracer genes in catecholaminergic neurons. Anat Cell Biol 43:157–164

    Article  PubMed  Google Scholar 

  12. He TC, Zhou SB, da Costa LT, Yu J, Kinzler KW, Vogelstein B (1998) A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 95:2509–2514

    Article  PubMed  CAS  Google Scholar 

  13. He, T. C. (2004) Adenoviral vectors, Current protocols in human genetics / editorial board, Jonathan L. Haines... [et al.] Chapter 12, Unit 12 14. John Wiley & Sons, Inc.

    Google Scholar 

  14. Luo J, Deng ZL, Luo X, Tang N, Song WX, Chen J, Sharff KA, Luu HH, Haydon RC, Kinzler KW, Vogelstein B, He TC (2007) A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. Nat Protoc 2:1236–1247

    Article  PubMed  CAS  Google Scholar 

  15. Luo Q, Kang Q, Song WX, Luu HH, Luo X, An N, Luo J, Deng ZL, Jiang W, Yin H, Chen J, Sharff KA, Tang N, Bennett E, Haydon RC, He TC (2007) Selection and validation of optimal siRNA target sites for RNAi-mediated gene silencing. Gene 395:160–169

    Article  PubMed  CAS  Google Scholar 

  16. Paxinos G, Franklin KBJ (2001) The mouse brain in stereotaxic coordinates, 2nd edn. Academic, New York, NY

    Google Scholar 

  17. Paxinos G, Watson C (2007) The rat brain in stereotaxic coordinates, 6th edn. Academic, San Diego, CA

    Google Scholar 

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Author information

Authors and Affiliations

  1. Division of Gastroenterology, Bayview Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Qian Li

  2. Molecular Oncology Laboratory, Department of Surgery, The University of Chicago Medical Center, Chicago, IL, USA

    Tong-Chuan He

Authors
  1. Qian Li
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  2. Tong-Chuan He
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Editor information

Editors and Affiliations

  1. Institute of Experimental Neurology, San Raffaele Scientific Institute Division of Neuroscience, Milan, Italy

    Riccardo Brambilla

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Li, Q., He, TC. (2014). Recombinant Adenovirus in Neurobiology. In: Brambilla, R. (eds) Viral Vector Approaches in Neurobiology and Brain Diseases. Neuromethods, vol 82. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-610-8_2

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  • DOI: https://doi.org/10.1007/978-1-62703-610-8_2

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-609-2

  • Online ISBN: 978-1-62703-610-8

  • eBook Packages: Springer Protocols

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