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BER Research Highlights


New Toolkit for Plant Genome Engineering
Published: June 01, 2017
Posted: August 02, 2021

Objective

Develop plant genome engineering techniques to allow efficient gene knockouts, replacements, altered transcriptional regulation, and multiplexed modifications.

Approach

  • A suite of direct and modular cloning vectors was developed to build TALEN-or CRISPR/Cas9-based editing targeting multiple genes and containing different promoters, reporters, selectable markers, and codon-optimized genes.
  • Vector construction is facilitated by an online tool that allows choice of different vector modules for different functions, either in monocot or dicot plants.

Results/Impact

  • The toolkit was tested for deletion mutagenesis targeting single or multiple genes, taking advantage of polycistronic transcripts with multiple guide RNAs (gRNA) in a single construct.
  • Up to 6 genes were successfully knocked out using multiplexed vectors, and a 58 Kbp genomic region containing 5 genes involved in nitrogen fixation was deleted in a single event.
  • The new toolkit will accelerate genome engineering of bioenergy crops, providing a flexible and comprehensive set of gene-editing options.

Cermák, T. et al. “Multipurpose toolkit to enable advanced genome engineering in plants.” The Plant Cell 29(6), 1196–1217 (2017). [DOI:10.1105/tpc.16.00922].

Topic Areas:

  • Research Area: Genomic Analysis and Systems Biology
  • Research Area: Plant Systems and Feedstocks, Plant-Microbe Interactions
  • Research Area: Biosystems Design

Division: SC-33.2 Biological Systems Science Division, BER

 

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