Research Team Combines Microspore Technology with CRISPR-Cas9May 9, 2018
The application of gene editing techniques such as TALENs and CRISPR-Cas9 is hindered by the long life cycle of crop species. Moreover, desired genotypes generally require multiple generations to achieve. Meanwhile, single-celled microspores are haploid cells that can be developed into double haploid plants and have been widely used as a breeding tool to generate homozygous plants within a generation.
A researcher from the National Research Council Canada, Pankaj Bhowmik, and his colleagues combined the CRISPR-Cas9 system with microspore technology to develop a haploid mutagenesis system to induce genetic modifications in the wheat genome.
The team first investigated the optimal conditions for microspore transfection of the CRISPR-Cas9 into microspores. Using multiple Cas9 and sgRNA constructs, the team successfully introduced targeted mutations in the reporter gene, DsRed, as well as two endogenous wheat genes, TaLox2 and TaUbiL1. However, the regeneration efficiency from microspores is still low and requires further study.
This study demonstrates the feasibility of combining microspore technology and CRISPR-Cas9 for trait discovery and crop improvement.
For more information, read the article in Scientific Reports.
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