Enabling Crop Trait Engineering, The Non-GMO Way
Threats of climate change and global pandemics continue to be factors that pressure our food and medicine security, increasing the need for more productive agriculture traits and biomanufacturing systems. Traditional crop breeding techniques are challenging and remain a lengthy process. Older generation transgenic technologies produce GMOs with permanent heterologous genes inserted, often with unknown effects.
Gene editing technology allows farmers to keep up with the ever increasing demands for more and better food, while using less water, land, nutrients and other resources. Advanced genetic engineering tools like Cas-CLOVER and piggyBac give plant biotechnologists the ability to make targeted modifications.
Scarless gene editing means no heterologous genes are left in the plant genome, producing a non-GMO crop and simplifying the production process. Making precise, intentional and beneficial changes in the genetic material of plants has the potential to optimize food production, as well as biofuels and therapeutic biomanufacturing.
Cas-CLOVER More Precise Than CRISPR
A clean alternative to CRISPR, Cas-CLOVER has the potential to revolutionize the world of agriculture by making precise, intentional and beneficial edits in the genetic material resulting in permanent changes.
A recent review identifies nearly 30 engineered traits for disease resistance and abiotic stress in important crops most of which will never be used on a commercial level due to CRISPR’s licensing restrictions.1Razzaq, A., Saleem, F., Kanwal, M., Mustafa, G., Yousaf, S., Arshad, H. M. I., … KhanJoyia, F. A. (2019). Modern trends in plant genome editing: An inclusive review of the CRISPR/Cas9 Toolbox. International Journal of Molecular Sciences, 20(16). https://doi.org/10.3390/ijms20164045
Like chemical mutagenesis but on a lower level, CRISPR-Cas9 also introduces abundant off-target mutations, increasing the need for backcrossing the crop to remove these unwanted mutations.
You can learn more about our more precise and easy-to-license Cas-CLOVER gene editing technology by clicking on the link below.
Cas-CLOVER Validated In Plants
We validated Cas-CLOVER in plants by efficiently creating knockout mutations in the model organism tobacco, followed by efficiency optimizations.
The phytoene desaturase (PDS) gene was targeted due to its visually pale and white phenotype. Green, pale mixed, and white shoots were all screened to determine cutting efficiencies. Afterwards, we verified that only white and pale material was edited at an overall average of around 50%+ editing efficiency (white shoots/total shoots from plates, upper image).
After fully optimizing our protocols (shared with you during evaluation) we enhanced both the number of white/pale shoots per plate and genomic cutting efficiencies up to 90%+ (lower image). See our blog post for more data including next-generation sequencing (NGS).
In fact, the vaccine for the last Ebola outbreak was produced in transgenic tobacco; showing the value of genetic editing tools in agriculture.
This research was presented at the Plant Biology 2020 virtual conference.
PiggyBac Validated In Rice For Trait Enhancement
As in most plants, homologous recombination (HR) mediated precision genome editing in rice is an infrequent event. Efficiency can be vastly improved by positive-negative selection. However, the positive selection marker (HPT) needs to be seamlessly removed from the edited locus.
piggyBac is the ideal technology for this task as it can be used for Footprint-Free excision following selection.
We offer licenses to piggyBac transposase for agricultural biotechnology.
Learn more about evaluating piggyBac through our easy-to-use licenses by clicking on the link below.
piggyBac Improves Herbicide Resistance In Rice
piggyBac-based gene editing was achieved at the rice ALS locus.
The targeting vector carries selection markers flanked by piggyBac target sequences and the desired mutations are located in the arms of homology.
Following selection of the edit and footprint-free excision, the ALS gene contains the desired point mutations with all other components of the targeting seamlessly removed.
The result is rice with point mutations resistant to the herbicide bispyribac sodium (BS).
Browse Gene Editing Reagents Online
Gene editing technologies have traditionally been accessible only through cost-prohibitive licensing terms that may require high upfront fees, milestones, and royalties.
We are providing simple license structures with accessible economic terms for Cas-CLOVER and piggyBac for agriculture biotechnology as well as proof-of-concept services.
To start, check out our easy-to-use catalog page. After requesting the vials and signing a simple royalty-free MTA form, we’ll send you our basic use protocol, which includes maps; sequences; instructions on design and production of the gRNAs.
Related Resources
Check out our latest content on how our gene editing technology is used in agriculture biotechnology.
See How Scientists Are Using Our Gene Editing Tools
Hear how Elanco Animal Health Research Scientist, Kayla Bean, Ph.D, explored how Cas-CLOVER optimizes the process for researchers who want to engineer the CHO and other genomes for cell line development aimed at protein expression.
“The flexibility of the guide RNA design makes the system easy to use and gives high specificity due to the use of the two guide RNAs.
It is very efficient due to the ability of the dead Cas9s to recognize the current area of DNA, and since the clo51 nuclease can only cut when dimerized, the system has high fidelity.”
Kayla Bean, Ph.D.
Research Scientist, Discovery Research
Elanco Animal Health
Easy-To License Gene Editing Tools
The validated Cas-CLOVER and piggyBac activity in plants is exciting for crop trait development as it opens up numerous opportunities.
We are offering clear commercial freedom to operate and simple accessible licenses to commercial users. We are also interested in special collaborations with academic groups that conduct research in traits which can ultimately be adopted commercially to enhance food production while being mindful of the environment.