GENE GUN ALLOWS TRANSIENT EXPRESSION OF CRISPR-CAS9 IN WHEAT
Wheat is recalcitrant to tissue culture, hindering its genetic improvement through transformation. Consequently, it cannot be improved through gene editing, which also requires transformation and tissue culture regeneration. To solve this problem, researcher Ryozo Imai from National Agriculture and Food Research Organization in Japan and colleagues use gene gun to transiently express CRISPR-Cas9 vector in wheat.
The researchers target the shoot apical meristem of mature wheat embryos for gene gun transformation. PCR screening was done to determine if the seeds were transformed. Afterwards, the seeds were germinated. Results show successful transformation of 5 percent of the seeds. Though low in efficiency, this method poses advantages as follows:
- Experiments can be done using mature embryos;
- Antibiotic selection is unnecessary; and
- RNA can also be introduced through a gene gun.
For more information, read the article in Scientific Reports.