We are developing a versatile method for efficiently editing plant DNA without adding DNA from other organisms.
Genome editing with the Cas9 protein-gRNA complex (RNPs) has tremendous potential for engineering plants because of its ability to induce genome editing without the use of DNA. DNA-free genome editing has the potential to significantly impact the field of agriculture, due to the rapid regulatory pathway associated with DNA-free genome editing procedures. However, delivering the Cas9 RNP into target plant tissues is challenging because of the presence of the cell wall, and the majority of drug delivery techniques developed for mammalian cells are ineffective on plant tissues. Biolistic delivery methods have great promise for genome editing in plants with the Cas9 RNP, given their widespread ability to deliver DNA into plants. However, the efficiency of biolistic Cas9 RNP delivery is currently low and strategies that can improve it have the potential to impact all areas of plant biology and agriculture. The central objective of this project is to develop a gold nanoparticle-Cas9 RNP complex, termed CRISPR-Gold, which can efficiently edit plant tissues after biolistic delivery.
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