Augmented sgRNAs for enhanced plant genome engineering (2019-322, Dr. Daniel Voytas)
The Voytas Lab has developed augmented sgRNAs that improve somatic cell genome editing and produce heritable modifications in plants using CRISPR/Cas9.
Plant gene editing requiring no regeneration and tissue culture
A new plant gene editing method improves somatic cell genome modifications and generates heritable genome modifications in plants. In addition, the method does not require regeneration and tissue culture. The novel method enables non-cell autonomous movement of sgRNA from the CRISPR/Cas RNA guided endonuclease system. It uses native plant mobile RNA and viral mobile RNA sequences to allow in-plant movement of editing reagents and enhance RNA guided endonuclease editing efficacy. First, an RNA viral vector expressing sgRNA fused to a mobile motif is introduced into a Cas9 expressing plant. Seeds collected from the infected plant create progeny with the desired mutation that requires no tissue culture. When combined with RNA viral vectors, this modified sgRNA gene editing method significantly enhances whole plant somatic cell gene editing in transgenic plants that overexpress the Cas9 endonuclease. This method also allows for these high percentage somatic gene edits to be transmitted to the next generation and become fixed in the germline.
Faster, efficient and less expensive
Despite remarkably rapid advances in the development of CRISPR/Cas9 technology and its many applications, it is still a challenge to perform genome engineering in plants. One of the major challenges in using CRISPR/Cas to make gene edited plants is transforming the plant with the gene editing reagents at high efficiency with an easy, inexpensive transformation method that results in heritable gene edits in the right cells. Currently, transforming plants is difficult and often requires tissue culture, which is both expensive and time consuming. This new method overcomes these limitations by using modified sgRNA sequences that increase somatic cell gene editing, allowing more efficient use of current gene editing and plant regeneration methods.
Phase of Development
- Proof of concept. Genome-editing approach has been validated in Nicotiana benthamiana.
Key Benefits & Differentiators
- Generates heritable genome modifications: Uses mobilized RNA elements in endonuclease-enabled genome editing to increase editing efficiency of somatic cells and facilitate delivery of editing reagents to meristematic tissues in plants.
- Reduces time, labor and costs: Bypasses the need for expensive and technically complex tissue culture and plant regeneration, resulting in genome-edited seed in a single generation.
- Applicable to a variety of plant species: Validated in dicots with work underway to demonstrate use in including agriculturally relevant species like tomato, wheat, maize and rice.
- Plant biotechnology
- Gene editing with CRISPR/Cas system
- Modified guide RNAs and viral vectors
- RNA guided endonucleases
- Site specific genome editing
- Gene knockouts
- Genetic structural modifications
- Sequence specific base editing
Ready for Licensing
This technology is now available for license! The University is excited to partner with industry to see this innovation reach its potential. Please contact us to share your business’ needs and your licensing interests in this technology. The license is for the sale, manufacture or use of products claimed by the patents.