Skip to main content

Distinct Expression Patterns of AAV8 Vectors with Broadly Active Promoters from Subretinal Injections of Neonatal Mouse Eyes at Two Different Ages

  • Conference paper
  • First Online:
Retinal Degenerative Diseases

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 854))

Abstract

The retinal expression patterns were analyzed following the injection of serotype 8 adeno-associated virus (AAV8) vectors that utilize two broadly active and commonly used sets of transcription regulatory sequences. These include the human cytomegalovirus (CMV) immediate early (IE) enhancer/promoter and the hybrid CAG element (also known as CAGGS or CBA) composed of a partial human CMV IE enhancer and the chicken β-actin promoter and intron. Subretinal delivery to postnatal day 0 (P0) or 6 (P6) mouse eyes resulted in efficient labeling of retinal cells, but with very distinct patterns. With P0 delivery, AAV8-CMV-GFP selectively labelled photoreceptors, while AAV8-CAG-GFP efficiently labeled both outer and inner retinal neurons, including photoreceptors, horizontal cells, amacrine cells and retinal ganglion cells. With P6 delivery, both vectors led to efficient labeling of photoreceptors and Müller glia cells, but not of inner retinal neurons. Our results suggest that the cell types that express the genes encoded by subretinally delivered AAV8 vectors are determined by both the timing of the injection and the regulatory sequences.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  • Allocca M, Mussolino C, Garcia-Hoyos M et al (2007) Novel adeno-associated virus serotypes efficiently transduce murine photoreceptors. J Virol 81:11372–11380

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Boshart M, Weber F, Jahn G et al (1985) A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41:521–530

    Article  CAS  PubMed  Google Scholar 

  • Cronin T, Vandenberghe LH, Hantz P et al (2014) Efficient transduction and optogenetic stimulation of retinal bipolar cells by a synthetic adeno-associated virus capsid and promoter. EMBO Mol Med 6(9):1175–1190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Doroudchi MM, Greenberg KP, Liu J et al (2011) Virally delivered channelrhodopsin-2 safely and effectively restores visual function in multiple mouse models of blindness. Mol Ther 19:1220–1229

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Macé E, Caplette R, Marre O et al (2015) Targeting channelrhodopsin-2 to ON-bipolar cells with vitreally administered AAV restores ON and OFF visual responses in blind mice. Mol Ther 23(1):7–16

    Article  PubMed  PubMed Central  Google Scholar 

  • Matsuda T, Cepko CL (2004) Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proc Natl Acad Sci USA 101:16–22

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McCarty DM, Young SM, Samulski RJ (2004) Integration of adeno-associated virus (AAV) and recombinant AAV vectors. Annu Rev Genet 38:819–845

    Article  CAS  PubMed  Google Scholar 

  • Niwa H, Yamamura K, Miyazaki J (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108:193–199

    Article  CAS  PubMed  Google Scholar 

  • Qin JY, Zhang L, Clift KL et al (2010) Systematic comparison of constitutive promoters and the doxycycline-inducible promoter. PLoS One 5:e10611

    Article  PubMed  PubMed Central  Google Scholar 

  • Uga S, Smelser GK (1973) Electron microscopic study of the development of retinal Müllerian cells. Invest Ophthalmol 12:295–307

    CAS  PubMed  Google Scholar 

  • Vandenberghe LH, Xiao R, Lock M et al (2010) Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing. Hum Gene Ther 21:1251–1257

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang S, Sengel C, Emerson M et al (2014) The binary decision of rod and bipolar fate in the vertebrate retina is controlled by a gene regulatory network. Dev Cell 30:513–527

    Google Scholar 

  • Watanabe S, Sanuki R, Ueno S et al (2013) Tropisms of AAV for subretinal delivery to the neonatal mouse retina and its application for in vivo rescue of developmental photoreceptor disorders. PLoS One 8:e54146

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Young RW (1985) Cell differentiation in the retina of the mouse. Anat Rec 212:199–205

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Constance Cepko .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer International Publishing Switzerland

About this paper

Cite this paper

Xiong, W., Cepko, C. (2016). Distinct Expression Patterns of AAV8 Vectors with Broadly Active Promoters from Subretinal Injections of Neonatal Mouse Eyes at Two Different Ages. In: Bowes Rickman, C., LaVail, M., Anderson, R., Grimm, C., Hollyfield, J., Ash, J. (eds) Retinal Degenerative Diseases. Advances in Experimental Medicine and Biology, vol 854. Springer, Cham. https://doi.org/10.1007/978-3-319-17121-0_67

Download citation

Publish with us

Policies and ethics