Skip to main content

Target-Induced SOFA-HDV Ribozyme

  • Protocol
  • First Online:
Ribozymes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 848))

Abstract

Small cis-acting ribozymes have been converted into trans-acting ribozymes possessing the ability to cleave RNA substrates. The Hepatitis Delta Virus (HDV) ribozyme is one of the rare examples of these that is derived from an RNA species that is found in human cells. Consequently, it possesses the natural ability to function in the presence of human proteins in addition to an outstanding stability in human cells, two significant advantages in its use. The development of an additional specific on/off adaptor (SOFA) has led to the production of a new generation of HDV ribozymes with improved specificities that provide a tool with significant potential for future development in the fields of both functional genomics and gene ­therapy. SOFA-HDV ribozyme-based gene inactivation systems have been reported in both prokaryotic and eukaryotic cells. Here, a step-by-step approach for the efficient design of highly specific SOFA-HDV ribozymes with a minimum investment of time and effort is described.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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

References

  1. Bagheri, S., and Kashani-Sabet, M. (2004) Ribozymes in the Age of Molecular Therapeutics. Curr. Mol. Med. 4, 489–506.

    Article  PubMed  CAS  Google Scholar 

  2. Schubert, S., and Kurreck, J. (2004) Ribozyme- and Deoxyribozyme-Strategies for Medical Applications. Curr. Drug Targets. 5, 667–681.

    Article  PubMed  CAS  Google Scholar 

  3. Asif-Ullah, M., Levesque, M., Robichaud, G., and Perreault, J. P. (2007) Development of Ribozyme-Based Gene-Inactivations; the Example of the Hepatitis Delta Virus Ribozyme. Curr. Gene Ther. 7, 205–216.

    Article  PubMed  CAS  Google Scholar 

  4. Tedeschi, L., Lande, C., Cecchettini, A., and Citti, L. (2009) Hammerhead Ribozymes in Therapeutic Target Discovery and Validation. Drug Discov. Today. 14, 776–783.

    Article  PubMed  CAS  Google Scholar 

  5. Teixeira, A., Tahiri-Alaoui, A., West, S., Thomas, B., Ramadass, A., Martianov, I., Dye, M., James, W., Proudfoot, N. J., and Akoulitchev, A. (2004) Autocatalytic RNA Cleavage in the Human Beta-Globin Pre-mRNA Promotes Transcription Termination. Nature. 432, 526–530.

    Article  PubMed  CAS  Google Scholar 

  6. Salehi-Ashtiani, K., Luptak, A., Litovchick, A., and Szostak, J. W. (2006) A Genomewide Search for Ribozymes Reveals an HDV-Like Sequence in the Human CPEB3 Gene. Science. 313, 1788–1792.

    Article  PubMed  CAS  Google Scholar 

  7. Levesque, D., Choufani, S., and Perreault, J. P. (2002) Delta Ribozyme Benefits from a Good Stability in Vitro that Becomes Outstanding in Vivo. RNA. 8, 464–477.

    Article  PubMed  CAS  Google Scholar 

  8. Bergeron, L. J., Ouellet, J., and Perreault, J. P. (2003) Ribozyme-Based Gene-Inactivation Systems Require a Fine Comprehension of their Substrate Specificities; the Case of Delta Ribozyme. Curr. Med. Chem. 10, 2589–2597.

    Article  PubMed  CAS  Google Scholar 

  9. Peracchi, A. (2004) Prospects for Antiviral Ribozymes and Deoxyribozymes. Rev. Med. Virol. 14, 47–64.

    Article  PubMed  CAS  Google Scholar 

  10. Bergeron, L. J., and Perreault, J. P. (2005) Target-Dependent on/off Switch Increases Ribozyme Fidelity. Nucleic Acids Res. 33, 1240–1248.

    Article  PubMed  CAS  Google Scholar 

  11. Bergeron, L. J., Reymond, C., and Perreault, J. P. (2005) Functional Characterization of the SOFA Delta Ribozyme. RNA. 11, 1858–1868.

    Article  PubMed  CAS  Google Scholar 

  12. Fiola, K., Perreault, J. P., and Cousineau, B. (2006) Gene Targeting in the Gram-Positive Bacterium Lactococcus lactis, using various Delta Ribozymes. Appl. Environ. Microbiol. 72, 869–879.

    Article  PubMed  CAS  Google Scholar 

  13. Bergeron, L. J., Reymond, C., and Perreault, J. P. (2005) Functional Characterization of the SOFA Delta Ribozyme. RNA. 11, 1858–1868.

    Article  PubMed  CAS  Google Scholar 

  14. Levesque, M. V., Levesque, D., Briere, F. P., and Perreault, J. P. (2010) Investigating a New Generation of Ribozymes in Order to Target HCV. PLoS One. 5, e9627.

    Article  PubMed  Google Scholar 

  15. Robichaud, G. A., Perreault, J. P., and Ouellette, R. J. (2008) Development of an Isoform-Specific Gene Suppression System: The Study of the Human Pax-5B Transcriptional Element. Nucleic Acids Res. 36, 4609–4620.

    Article  PubMed  CAS  Google Scholar 

  16. von Eije, K. J., ter Brake, O., and Berkhout, B. (2008) Human Immunodeficiency Virus Type 1 Escape is Restricted when Conserved Genome Sequences are Targeted by RNA Interference. J. Virol. 82, 2895–2903.

    Article  Google Scholar 

  17. Haasnoot, J., Westerhout, E. M., and Berkhout, B. (2007) RNA Interference Against Viruses: Strike and Counterstrike. Nat. Biotechnol. 25, 1435–1443.

    Article  PubMed  CAS  Google Scholar 

  18. Amarzguioui, M., Brede, G., Babaie, E., Grotli, M., Sproat, B., and Prydz, H. (2000) Secondary Structure Prediction and in Vitro Accessibility of mRNA as Tools in the Selection of Target Sites for Ribozymes. Nucleic Acids Res. 28, 4113–4124.

    Article  PubMed  CAS  Google Scholar 

  19. Ryu, K. J., and Lee, S. W. (2004) Comparative Analysis of Intracellular Trans-Splicing Ribozyme Activity Against Hepatitis C Virus Internal Ribosome Entry Site. J. Microbiol. 42, 361–364.

    PubMed  CAS  Google Scholar 

  20. Doran, G., and Sohail, M. (2006) Systematic Analysis of the Role of Target Site Accessibility in the Activity of DNA Enzymes. J. RNAi Gene Silencing. 2, 205–214.

    PubMed  CAS  Google Scholar 

  21. Deschenes, P., Lafontaine, D. A., Charland, S., and Perreault, J. P. (2000) Nucleotides -1 to -4 of Hepatitis Delta Ribozyme Substrate Increase the Specificity of Ribozyme Cleavage. Antisense Nucleic Acid Drug Dev. 10, 53–61.

    Article  PubMed  CAS  Google Scholar 

  22. Lucier, J. F., Bergeron, L. J., Briere, F. P., Ouellette, R., Elela, S. A., and Perreault, J. P. (2006) RiboSubstrates: A Web Application Addressing the Cleavage Specificities of Ribozymes in Designated Genomes. BMC Bioinformatics. 7, 480.

    Article  PubMed  Google Scholar 

  23. Ouellet, J., and Perreault, J. P. (2004) Cross-Linking Experiments Reveal the Presence of Novel Structural Features between a Hepatitis Delta Virus Ribozyme and its Substrate. RNA. 10, 1059–1072.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Jonathan Perreault and Gilles Boire for information about hY RNA. This work was supported by grants from Canadian Institute of Health Research (CIHR; grant numbers MOP-44002 and EOP-38322) to J.P.P. The RNA group is supported by grants both from CIHR and Université de Sherbrooke. M.V.L. was the recipient of a predoctoral fellowship from the Fonds de Recherche en Santé du Québec. J.P.P. holds the Canada Research Chairs in genomics and catalytic RNAs, and is a member of the Centre de Recherche Clinique Étienne Lebel.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jean-Pierre Perreault .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Lévesque, M.V., Perreault, JP. (2012). Target-Induced SOFA-HDV Ribozyme. In: Hartig, J. (eds) Ribozymes. Methods in Molecular Biology, vol 848. Humana Press. https://doi.org/10.1007/978-1-61779-545-9_23

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-545-9_23

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-544-2

  • Online ISBN: 978-1-61779-545-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics