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The heat is on : a simple method to increase genome editing efficiency in plants

Jonas Blomme (UGent) , Ward Develtere (UGent) , Ayse Köse, Júlia Arraiza (UGent) , Christophe Brugmans, Jessica Jaraba Wallace, Ward Decaestecker, Debbie Rombaut (UGent) , Alexandra Baekelandt (UGent) , Álvaro Daniel Fernández Fernández, et al.
(2022) BMC PLANT BIOLOGY. 22(1).
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Abstract
Background Precision genome mutagenesis using CRISPR/Cas has become the standard method to generate mutant plant lines. Several improvements have been made to increase mutagenesis efficiency, either through vector optimisation or the application of heat stress. Results Here, we present a simplified heat stress assay that can be completed in six days using commonly-available laboratory equipment. We show that three heat shocks (3xHS) efficiently increases indel efficiency of LbCas12a and Cas9, irrespective of the target sequence or the promoter used to express the nuclease. The generated indels are primarily somatic, but for three out of five targets we demonstrate that up to 25% more biallelic mutations are transmitted to the progeny when heat is applied compared to non-heat controls. We also applied our heat treatment to lines containing CRISPR base editors and observed a 22-27% increase in the percentage of C-to-T base editing. Furthermore, we test the effect of 3xHS on generating large deletions and a homologous recombination reporter. Interestingly, we observed no positive effect of 3xHS treatment on either approach using our conditions. Conclusions Together, our experiments show that heat treatment is consistently effective at increasing the number of somatic mutations using many CRISPR approaches in plants and in some cases can increase the recovery of mutant progeny.
Keywords
Base editing, CRISPR, Genome engineering, Heat-shock, Homology-directed repair, Plant biotechnology, ARABIDOPSIS, MUTAGENESIS, CRISPR/CAS9, EXPRESSION, NUCLEASES, GENE, DNA, RECOMBINATION, ENDONUCLEASE, TEMPERATURE

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MLA
Blomme, Jonas, et al. “The Heat Is on : A Simple Method to Increase Genome Editing Efficiency in Plants.” BMC PLANT BIOLOGY, vol. 22, no. 1, 2022, doi:10.1186/s12870-022-03519-7.
APA
Blomme, J., Develtere, W., Köse, A., Arraiza, J., Brugmans, C., Jaraba Wallace, J., … Jacobs, T. B. (2022). The heat is on : a simple method to increase genome editing efficiency in plants. BMC PLANT BIOLOGY, 22(1). https://doi.org/10.1186/s12870-022-03519-7
Chicago author-date
Blomme, Jonas, Ward Develtere, Ayse Köse, Júlia Arraiza, Christophe Brugmans, Jessica Jaraba Wallace, Ward Decaestecker, et al. 2022. “The Heat Is on : A Simple Method to Increase Genome Editing Efficiency in Plants.” BMC PLANT BIOLOGY 22 (1). https://doi.org/10.1186/s12870-022-03519-7.
Chicago author-date (all authors)
Blomme, Jonas, Ward Develtere, Ayse Köse, Júlia Arraiza, Christophe Brugmans, Jessica Jaraba Wallace, Ward Decaestecker, Debbie Rombaut, Alexandra Baekelandt, Álvaro Daniel Fernández Fernández, Frank Van Breusegem, Dirk Inzé, and Thomas B. Jacobs. 2022. “The Heat Is on : A Simple Method to Increase Genome Editing Efficiency in Plants.” BMC PLANT BIOLOGY 22 (1). doi:10.1186/s12870-022-03519-7.
Vancouver
1.
Blomme J, Develtere W, Köse A, Arraiza J, Brugmans C, Jaraba Wallace J, et al. The heat is on : a simple method to increase genome editing efficiency in plants. BMC PLANT BIOLOGY. 2022;22(1).
IEEE
[1]
J. Blomme et al., “The heat is on : a simple method to increase genome editing efficiency in plants,” BMC PLANT BIOLOGY, vol. 22, no. 1, 2022.
@article{8748416,
  abstract     = {{Background
Precision genome mutagenesis using CRISPR/Cas has become the standard method to generate mutant plant lines. Several improvements have been made to increase mutagenesis efficiency, either through vector optimisation or the application of heat stress.

Results
Here, we present a simplified heat stress assay that can be completed in six days using commonly-available laboratory equipment. We show that three heat shocks (3xHS) efficiently increases indel efficiency of LbCas12a and Cas9, irrespective of the target sequence or the promoter used to express the nuclease. The generated indels are primarily somatic, but for three out of five targets we demonstrate that up to 25% more biallelic mutations are transmitted to the progeny when heat is applied compared to non-heat controls. We also applied our heat treatment to lines containing CRISPR base editors and observed a 22-27% increase in the percentage of C-to-T base editing. Furthermore, we test the effect of 3xHS on generating large deletions and a homologous recombination reporter. Interestingly, we observed no positive effect of 3xHS treatment on either approach using our conditions.

Conclusions
Together, our experiments show that heat treatment is consistently effective at increasing the number of somatic mutations using many CRISPR approaches in plants and in some cases can increase the recovery of mutant progeny.}},
  articleno    = {{142}},
  author       = {{Blomme, Jonas and Develtere, Ward and Köse, Ayse and Arraiza, Júlia and Brugmans, Christophe and Jaraba Wallace, Jessica and Decaestecker, Ward and Rombaut, Debbie and Baekelandt, Alexandra and Fernández Fernández, Álvaro Daniel and Van Breusegem, Frank and Inzé, Dirk and Jacobs, Thomas B.}},
  issn         = {{1471-2229}},
  journal      = {{BMC PLANT BIOLOGY}},
  keywords     = {{Base editing,CRISPR,Genome engineering,Heat-shock,Homology-directed repair,Plant biotechnology,ARABIDOPSIS,MUTAGENESIS,CRISPR/CAS9,EXPRESSION,NUCLEASES,GENE,DNA,RECOMBINATION,ENDONUCLEASE,TEMPERATURE}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{14}},
  title        = {{The heat is on : a simple method to increase genome editing efficiency in plants}},
  url          = {{http://doi.org/10.1186/s12870-022-03519-7}},
  volume       = {{22}},
  year         = {{2022}},
}
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