Skip to main content

Computational Detection of CRISPR/crRNA Targets

  • Protocol
CRISPR

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1311))

Abstract

The CRISPR-Cas systems in bacteria and archaea provide protection by targeting foreign nucleic acids. The sequence of the “spacers” within CRISPR arrays specifically determines the targets in invader genomes. These spacers provide the short specific RNA nucleotide sequences within the guide crRNAs. In addition to complementarity in the spacer–target (protospacer) interaction, short flanking protospacer adjacent motifs (PAMs), or mismatching flanks have a discriminatory role in accurate target detection. Here, we describe a bioinformatic method, called CRISPRTarget, to use the sequence of a CRISPR array (e.g., predicted via CRISPRDetect/CRISPRDirection) to identify the foreign nucleic acids it targets.

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 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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. Richter C, Chang JT, Fineran PC (2012) Function and regulation of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) systems. Viruses 4:2291–2311

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Westra ER, Buckling A, Fineran PC (2014) CRISPR-Cas systems: beyond adaptive immunity. Nat Rev Microbiol 12:317–326

    Article  CAS  PubMed  Google Scholar 

  3. Drevet C, Pourcel C (2012) How to identify CRISPRs in sequencing data. Methods Mol Biol 905:15–27

    CAS  PubMed  Google Scholar 

  4. Lange SJ, Alkhnbashi OS, Rose D, Will S, Backofen R (2013) CRISPRmap: an automated classification of repeat conservation in prokaryotic adaptive immune systems. Nucleic Acids Res 41:8034–8044

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Biswas A, Fineran PC, Brown CM (2014) Accurate computational prediction of the transcribed strand of CRISPR non-coding RNAs. Bioinformatics 30:1805–1813

    Article  CAS  PubMed  Google Scholar 

  6. Biswas A, Gagnon JN, Brouns SJ, Fineran PC, Brown CM (2013) CRISPRTarget: bioinformatic prediction and analysis of crRNA targets. RNA Biol 10:817–827

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Fineran PC, Gerritzen MJ, Suarez-Diez M, Kunne T, Boekhorst J, van Hijum SA, Staals RH, Brouns SJ (2014) Degenerate target sites mediate rapid primed CRISPR adaptation. Proc Natl Acad Sci U S A 111:E1629–E1638

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Richter C, Dy RL, McKenzie RE, Watson BN, Taylor C, Chang JT, McNeil M, Staals RHJ, Fineran PC (2014) Priming in the Type I-F CRISPR-Cas system triggers strand-independent spacer acquisition, bi-directionally from the primed protospacer. Nucleic Acids Res 42(13):8516–8526

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero DA, Horvath P (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315:1709–1712

    Article  CAS  PubMed  Google Scholar 

  10. Deveau H, Barrangou R, Garneau JE, Labonte J, Fremaux C, Boyaval P, Romero DA, Horvath P, Moineau S (2008) Phage response to CRISPR-encoded resistance in Streptococcus thermophilus. J Bacteriol 190:1390–1400

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Nat Protoc 8:2281–2308

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Fineran PC, Dy RL (2014) Gene regulation by engineered CRISPR-Cas systems. Curr Opin Microbiol 18:83–89

    Article  CAS  PubMed  Google Scholar 

  13. Yang L, Mali P, Kim-Kiselak C, Church G (2014) CRISPR-Cas-mediated targeted genome editing in human cells. Methods Mol Biol 1114:245–267

    Article  CAS  PubMed  Google Scholar 

  14. Bland C, Ramsey TL, Sabree F, Lowe M, Brown K, Kyrpides NC, Hugenholtz P (2007) CRISPR recognition tool (CRT): a tool for automatic detection of clustered regularly interspaced palindromic repeats. BMC Bioinformatics 8:209

    Article  PubMed Central  PubMed  Google Scholar 

  15. Edgar RC (2007) PILER-CR: fast and accurate identification of CRISPR repeats. BMC Bioinformatics 8:18

    Article  PubMed Central  PubMed  Google Scholar 

  16. Grissa I, Vergnaud G, Pourcel C (2007) CRISPRFinder: a web tool to identify clustered regularly interspaced short palindromic repeats. Nucleic Acids Res 35:W52–W57

    Article  PubMed Central  PubMed  Google Scholar 

  17. Rousseau C, Gonnet M, Le Romancer M, Nicolas J (2009) CRISPI: a CRISPR interactive database. Bioinformatics 25:3317–3318

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Skennerton CT, Imelfort M, Tyson GW (2013) Crass: identification and reconstruction of CRISPR from unassembled metagenomic data. Nucleic Acids Res 41:e105

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Rho M, Wu YW, Tang H, Doak TG, Ye Y (2012) Diverse CRISPRs evolving in human microbiomes. PLoS Genet 8:e1002441

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by a Rutherford Discovery Fellowship from the Royal Society of NZ to PCF, by a Human Frontier Science Program Grant to Ian Macara, Anne Spang and CMB. AB was a recipient of a University of Otago Postgraduate Scholarship and a Postgraduate Publishing Bursary.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Chris M. Brown .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Biswas, A., Fineran, P.C., Brown, C.M. (2015). Computational Detection of CRISPR/crRNA Targets. In: Lundgren, M., Charpentier, E., Fineran, P. (eds) CRISPR. Methods in Molecular Biology, vol 1311. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2687-9_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-2687-9_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2686-2

  • Online ISBN: 978-1-4939-2687-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics