Skip to main content

Assembly of TALEN and mTALE-Act for Plant Genome Engineering

  • Protocol
  • First Online:
Crop Breeding

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2264))

Abstract

Transcription activator-like effector (TALE) is a DNA-binding domain that can be paired with a nuclease to create DNA double-strand breaks, or with an effector protein to alter gene transcription. The ability to precisely alter plant genomes and transcriptomes has provided many insights into gene function and has recently been utilized for crop improvement. Easy design and construction of TALE make the tool more accessible to a variety of researchers. Here, we describe two TALE-based systems: transcription activator-like effector nucleases (TALEN), for creating targeted mutations in a gene of interest, and multiplex TALE activation (mTALE-Act), for activating one or a few genes of interest at the transcription level. Assembly of these tools is based on Golden Gate cloning and Gateway recombination, which are cost-effective and streamlined cloning methods.

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

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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 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

  1. Paul JW, Qi Y (2016) CRISPR/Cas9 for plant genome editing: accomplishments, problems and prospects. Plant Cell Rep 35:1417–1427. https://doi.org/10.1007/s00299-016-1985-z

    Article  CAS  PubMed  Google Scholar 

  2. Malzahn A, Lowder L, Qi Y (2017) Plant genome editing with TALEN and CRISPR. Cell Biosci 7. https://doi.org/10.1186/s13578-017-0148-4

  3. Römer P, Hahn S, Jordan T, Strauß T, Bonas U, Lahaye T (2007) Plant pathogen recognition mediated by promoter activation of the pepper Bs3 resistance gene. Science 318:645–648. https://doi.org/10.1126/science.1144958

    Article  CAS  PubMed  Google Scholar 

  4. Boch J, Scholze H, Schornack S, Landgraf A, Hahn S, Kay S, Lahaye T, Nickstadt A, Bonas U (2009) Breaking the code of DNA binding specificity of TAL-type III effectors. Science 326:1509–1512. https://doi.org/10.1126/science.1178811

    Article  CAS  PubMed  Google Scholar 

  5. Moscou MJ, Bogdanove AJ (2009) A simple cipher governs DNA recognition by TAL effectors. Science 326:1501. https://doi.org/10.1126/science.1178817

    Article  CAS  PubMed  Google Scholar 

  6. Christian M, Cermak T, Doyle EL, Schmidt C, Zhang F, Hummel A, Bogdanove AJ, Voytas DF (2010) Targeting DNA double-strand breaks with TAL effector nucleases. Genetics 186:757–761. https://doi.org/10.1534/genetics.110.120717

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Zhang Y, Zhang F, Li X, Baller JA, Qi Y, Starker CG, Bogdanove AJ, Voytas DF (2013) Transcription activator-like effector nucleases enable efficient plant genome engineering. Plant Physiol 161:20–27. https://doi.org/10.1104/pp.112.205179

    Article  CAS  PubMed  Google Scholar 

  8. Christian M, Qi Y, Zhang Y, Voytas DF (2013) Targeted mutagenesis of Arabidopsis thaliana using engineered TAL effector nucleases. G3 3:1697–1705. https://doi.org/10.1534/g3.113.007104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Li T, Huang S, Jiang WZ, Wright D, Spalding MH, Weeks DP, Yang B (2011) TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain. Nucleic Acids Res 39:359–372. https://doi.org/10.1093/nar/gkq704

    Article  CAS  PubMed  Google Scholar 

  10. Lowder LG, Zhou J, Zhang Y, Malzahn A, Zhong Z, Hsieh T-F, Voytas DF, Zhang Y, Qi Y (2018) Robust transcriptional activation in plants using multiplexed CRISPR-Act2.0 and mTALE-act systems. Mol Plant 11:245–256. https://doi.org/10.1016/j.molp.2017.11.010

    Article  CAS  PubMed  Google Scholar 

  11. Gammage PA, Moraes CT, Minczuk M (2018) Mitochondrial genome engineering: the revolution may not be CRISPR-Ized. Trends Genet 34:101–110. https://doi.org/10.1016/j.tig.2017.11.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF (2011) Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res 39:e82. https://doi.org/10.1093/nar/gkr218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Doyle EL, Booher NJ, Standage DS, Voytas DF, Brendel VP, VanDyk JK, Bogdanove AJ (2012) TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for TAL effector design and target prediction. Nucleic Acids Res 40:W117–W122. https://doi.org/10.1093/nar/gks608

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Streubel J, Blücher C, Landgraf A, Boch J (2012) TAL effector RVD specificities and efficiencies. Nat Biotechnol 30:593–595. https://doi.org/10.1038/nbt.2304

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work is supported by NSF Plant Genome ECA-PGR Award (IOS-1758745) to the University of Maryland, College Park.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yiping Qi .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Malzahn, A.A., Qi, Y. (2021). Assembly of TALEN and mTALE-Act for Plant Genome Engineering. In: Tripodi, P. (eds) Crop Breeding. Methods in Molecular Biology, vol 2264. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1201-9_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1201-9_15

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1200-2

  • Online ISBN: 978-1-0716-1201-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics