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Creating designer plants using CRISPR technology: A non-GMO approach

Principal Supervisor: Stephen Jackson, School of Life Sciences and Yiguo Hong, Hangzhou Univ., China

PhD project title: Creating designer plants using CRISPR technology: A non-GMO approach

University of Registration: Warwick

Project outline:

For many years it has been possible to manipulate aspects of plant development through the creation of transgenic plants expressing exogenous genes, however it has always been a long-term goal to do this by altering endogenous genes without the need for transgenic plants. Recently technologies have been developed that now enable gene-editing of endogenous genes, such techniques include Zinc finger nucleases, TALENs, and the more recent CRISPR RNA gene editing system (Cong et al. 2013, de Souza 2013). We will investigate the possibility of using the CRISPR system, to alter the sequences of genes involved in the control of flowering in order to manipulate the flowering time of crop plants.

We have a virus-based expression system which we have already used successfully to express functional proteins in tobacco plants (Li et al. 2009). We will test whether this expression system can be used to deliver the gene-editing proteins/RNA into plant cells and to create gene-edited plants. Other approaches that have already been published such as expression in plant protoplasts followed by regeneration of gene-edited plants will also be tested. In this project we will target genes involved in controlling flowering time in crop plants such as FLOWERING LOCUS T (FT) and FLC. If such approaches are successful then this will revolutionize the way in which genetically modified plants are produced, enabling them to be produced without the need to incorporate any foreign DNA into their genome.

References:

  • Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., ... & Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science, 339(6121), 819-823.
  • de Souza, N. (2013). Genetics: RNA-guided gene editing. Nature Methods, 10(3), 189-189.
  • Li, C., Zhang, K., Zeng ,X., Jackson, S., Zhou, Y., and Y. Hong (2009). A cis-element within FLOWERING LOCUS T mRNA determines its mobility and facilitates trafficking of heterologous viral RNA. J.Virology 83:3540-3548

BBSRC Strategic Research Priority: Food security

Techniques that will be undertaken during the project:

  • Molecular biology techniques such as cloning and gene expression using real-time PCR
  • Sequencing and analysis
  • Gene editing using CRISPR technology
  • Plant cell tissue culture and transformation
  • Plant physiology

Contact: Dr Stephen Jackson, University of Warwick