The Idea
The core thought behind ENABLE® builds on the scientific expertise of our Grow More team to translate plant science technologies to underutilized crops. We want to promote equitable access to science and solve global agricultural challenges by providing ENABLE® Kits, resources, and scientific insights.
Our first-generation ENABLE® Gene Editing in planta kits utilize the most advanced and precise gene editing method to date: CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats). They enable the user to generate targeted deletions and knockouts in their crop gene(s) of interest by providing the necessary plasmids to generate CRISPR constructs ready for plant transformation alongside extensive documentation and protocols. Please read all about the toolkit creation, its components and the in planta testing that we performed in our corresponding preprint.
Currently, the kit is distributed via Addgene. If you are from a low and middle income country and and have problems obtaining the vectors from Addgene, please get in touch with us!
The ENABLE® Gene Editing Kit contents
To induce a mutation in a gene using CRISPR/Cas9, plants are generally transformed with a vector containing the genes for Cas9, sgRNA(s) and a selection marker for verification of succesful transformation. We provide a vector suite containing vectors for sgRNA expression (pGMF1 and pGMF2) for monocots or dicots, a vector for Cas9 expression (pGMF4), two different selection markers (pGMF5 and pGMF6) and a binary T-DNA backbone (pGFM3) suitable for stable or transient transformation methods. The user can combine those components using the well established and simple Golden Gate cloning method into a single CRISPR vector for plant transformation and modification of any gene of choice. You can order all vectors at Addgene.
Extensive documentation and support
We want to enable all kit users to become proficient in gene editing and to fully understand each step of the process. That is why we provide extensive documentation along with the vectors which will support the user in crucial parts of the project, such as CRISPR target site choice and the Golden Gate cloning procedure. We understand that every project can face unique challenges and therefore also offer a free consulting service alongside our kits. Please do not hesitate to discuss your projects with us and provide us with valuable feedback about our CRISPR kits which we can incorperate into future releases.
The following protocols have been developed for new and experienced users of the ENABLE Gene Editing in planta toolkit:
Extended protocol for new users
Prerequisites
We are happy to provide our ENABLE® kits via Addgene to every active academic researcher in low and middle income countries and beyond. While we provide all the relevant vectors for a CRISPR project, we can neither provide enzymes and reagents, nor general molecular biology or plant transformation reagents at this stage. Please ensure that you have access to standard molecular biology equipment (e.g. pipettes, PCR tubes, thermocycler, Miniprep reagents), Oligo DNA/primer suppliers (you will need to order four DNA oligos to clone your unique CRISPR target sites) and methods to verify integrity of the cloned CRISPR vector (Sanger sequencing services, PCR based methods or restriction digest based methods). Similarly, reagents for E. coli, A. tumefaciens, or plant/protoplast transformation will need to be supplied by the user. If you need support with any of those things, please get in touch and we will try to find a solution together.
