Skip to main content
SpringerLink
Log in
Book cover

Morpholino Oligomers pp 215–227Cite as

In Vivo and Explant Electroporation of Morpholinos in the Developing Mouse Retina

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1565))

Abstract

Neonatal in vivo electroporations and retinal explant electroporations have been widely employed in understanding the effects of loss or gain of function of protein-coding genes in retinal development. Here, we describe a rapid and efficient delivery of morpholinos to add another tool to perturb gene expression during mouse retinal development.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   159.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Kaida D, Berg MG, Younis I, Kasim M, Singh LN, Wan L, Dreyfuss G (2010) U1 snRNP protects pre-mRNAs from premature cleavage and polyadenylation. Nature 468(7324):664–668. doi:10.1038/nature09479

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Sasaki Y, Welshhans K, Wen Z, Yao J, Xu M, Goshima Y, Zheng JQ, Bassell GJ (2010) Phosphorylation of zipcode binding protein is required for BDNF signaling of local β-actin synthesis and growth cone turning. J Neurosci 30(28):9349–9358

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Eom T, Antar LN, Singer RH, Bassell GJ (2003) Localization of a beta-actin messenger ribonucleoprotein complex with zipcode-binding protein modulates the density of dendritic filopodia and filopodial synapses. J Neurosci 23(32):10433–10444

    CAS  PubMed  Google Scholar 

  4. Staton AA, Giraldez AJ (2011) Use of target protector morpholinos to analyze the physiological roles of specific miRNA-mRNA pairs in vivo. Nat Protoc 6(12):2035–2049

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Cornell RA, Eisen JS (2000) Delta signaling mediates segregation of neural crest and spinal sensory neurons from zebrafish lateral neural plate. Development 127(13):2873–2882

    CAS  PubMed  Google Scholar 

  6. Cornell RA, Eisen JS (2002) Delta/Notch signaling promotes formation of zebrafish neural crest by repressing Neurogenin 1 function. Development 129(11):2639–2648

    CAS  PubMed  Google Scholar 

  7. Knapp D, Messenger N, Ahmed Rana A, Smith JC (2006) Neurotrophin receptor homolog (NRH1) proteins regulate mesoderm formation and apoptosis during early Xenopus development. Dev Biol 300(2):554–569. doi:10.1016/j.ydbio.2006.09.038

    Article  CAS  PubMed  Google Scholar 

  8. Flynt AS, Li N, Thatcher EJ, Solnica-Krezel L, Patton JG (2007) Zebrafish miR-214 modulates Hedgehog signaling to specify muscle cell fate. Nat Genet 39(2):259–263. doi:10.1038/ng1953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Eberhart JK, He X, Swartz ME, Yan YL, Song H, Boling TC, Kunerth AK, Walker MB, Kimmel CB, Postlethwait JH (2008) MicroRNA Mirn140 modulates Pdgf signaling during palatogenesis. Nat Genet 40(3):290–298. doi:10.1038/ng.82

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Martello G, Zacchigna L, Inui M, Montagner M, Adorno M, Mamidi A, Morsut L, Soligo S, Tran U, Dupont S, Cordenonsi M, Wessely O, Piccolo S (2007) MicroRNA control of Nodal signalling. Nature 449(7159):183–188. doi:10.1038/nature06100

    Article  CAS  PubMed  Google Scholar 

  11. Pekarik V, Bourikas D, Miglino N, Joset P, Preiswerk S, Stoeckli ET (2003) Screening for gene function in chicken embryo using RNAi and electroporation. Nat Biotechnol 21(1):93–96. doi:10.1038/nbt770

    Article  CAS  PubMed  Google Scholar 

  12. Muramatsu T (2000) In ovo gene electroporation into early chicken embryos. Methods Mol Med 37:369–376. doi:10.1385/1-59259-080-2:369

    CAS  PubMed  Google Scholar 

  13. Saito T, Nakatsuji N (2001) Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev Biol 240(1):237–246. doi:10.1006/dbio.2001.0439

    Article  CAS  PubMed  Google Scholar 

  14. Livesey FJ, Cepko CL (2001) Vertebrate neural cell-fate determination: lessons from the retina. Nat Rev Neurosci 2(2):109–118. doi:10.1038/35053522

    Article  CAS  PubMed  Google Scholar 

  15. Cepko CL, Austin CP, Yang X, Alexiades M, Ezzeddine D (1996) Cell fate determination in the vertebrate retina. Proc Natl Acad Sci U S A 93(2):589–595

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Matsuda T, Cepko CL (2004) Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proc Natl Acad Sci U S A 101(1):16–22. doi:10.1073/pnas.2235688100

    Article  CAS  PubMed  Google Scholar 

  17. Matsuda T, Cepko CL (2008) Analysis of gene function in the retina. Methods Mol Biol 423:259–278. doi:10.1007/978-1-59745-194-9_19

    Article  CAS  PubMed  Google Scholar 

  18. Barnabe-Heider F, Meletis K, Eriksson M, Bergmann O, Sabelstrom H, Harvey MA, Mikkers H, Frisen J (2008) Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation. Nat Methods 5(2):189–196. doi:10.1038/nmeth.1174

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by P30 from National Institute of Neurological Disorders and Stroke (5P30NS069266) and K99-R00 from National Eye Institute (4R00EY019547) to R.K. We would like to thank Edward J. Lechowicz, Jr. at the University of Connecticut for building the explant electroporation chamber.

Author information

Authors and Affiliations

  1. Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Road, Storrs, CT, 06269, USA

    Devi Krishna Priya Karunakaran & Rahul Kanadia

Authors
  1. Devi Krishna Priya Karunakaran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rahul Kanadia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rahul Kanadia .

Editor information

Editors and Affiliations

  1. Department of Biomedical Sciences College of Veterinary Medicine, Oregon State University, Corvallis, Oregon, USA

    Hong M. Moulton

  2. Gene Tools, LLC, Philomath, Oregon, USA

    Jon D. Moulton

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Karunakaran, D.K.P., Kanadia, R. (2017). In Vivo and Explant Electroporation of Morpholinos in the Developing Mouse Retina. 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_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6817-6_18

  • 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

Publish with us

Policies and ethics

Search

Navigation