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ENABLE® Gene Editing in planta Toolkit
(Kit # 1000000270 )

Depositing Lab:   Kate Creasey Krainer, Vladimir Nekrasov, Nicola Patron

The ENABLE® Gene Editing in planta toolkit provides a streamlined set of molecular tools for constructing CRISPR gene-editing vectors in just two simple cloning steps. Compatible with both monocot and dicot species, the kit supports transient and stable transformation approaches and is designed for use by researchers with any level of CRISPR experience.

This kit will be sent as individual bacterial stabs at room temperature.

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$385 USD + shipping
Available to academics and nonprofits only.
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Original Publication

Developing a Molecular Toolkit to ENABLE all to apply CRISPR/Cas9-based Gene Editing in planta. Abate BA, Hahn F, Chirivì D, Betti C, Fornara F, Molloy JC, Creasey Krainer KM. bioRxiv. 2025 November 13. doi: 10.1101/2025.11.09.687425. Article (Link opens in a new window)

Description

The Grow More Foundation (Link opens in a new window) is a nonprofit organization dedicated to ensuring equitable access to plant biotechnology in low- and middle-income countries. Through its ENABLE® program, the foundation works to turn advanced plant science into practical solutions for underutilized staple crops, empowering scientists in the Global South to address the agricultural challenges most relevant to their communities.

Our goal is to reduce technical and financial barriers to gene editing, allowing students, professors, researchers, and others to use genome-editing tools in local research projects. We see ENABLE® as a foundation for hands-on training, innovation, and creating new traits in neglected and underused crops — and as a step toward making crop research and development more accessible worldwide.

We created the ENABLE® Gene Editing in planta toolkit, a straightforward molecular toolkit that includes eight CRISPR-related plasmids along with comprehensive documentation. Our toolkit enables gene editing in both monocot and dicot plants and is suitable for stable or transient plant transformation.

For more detail, read the accompanying publication (Abate et al., 2025 (Link opens in a new window)) and protocols linked in the Protocols & Resources tab, or visit the Grow More Foundation's resource page (Link opens in a new window).

The schematic is shown as a vertical branching tree diagram with four levels. The top level shows the eight plasmids in the ENABLE gene editing in plant toolkit: pGMF1-M containing gRNA1, pGMF2-M containing gRNA2, pGMF1-D containing gRNA1, pGMF2-D containing gRNA2, pGMF3 (plasmid backbone), pGMF4 (pFH54) containing Cas9, pGMF5 (pICSL11059) containing hygromycin resistance, and pGMF6 containing eGFP. The second layer branches into plant species, with monocots to the left and dicots to the right. The monocots branch contains pGMF1-M and pGMF2-M, with gRNAs 1 and 2, respectively. The dicots branch contains pGMF1-D and pGMF2-D, with gRNAs 1 and 2, respectively. For the third level, monocot and dicot each split based on transformation type, with stable on the left and transient on the right. The stable branch for both monocot and dicot contains pGMF3, pGMF4, and pGMF5. The transient branch for both monocot and dicot contains pGMF3, pGMF4, and pGMF6. The fourth branch layer shows cartoon schematics of the outcome, with the stable transformation branches showing plants sprouting in a petri dish, and the transient showing cells fluorescing under a microscope.
Figure 1: Eight plasmids are provided to the user as part of the ENABLE® Gene Editing in planta toolkit. The user selects five of these plasmids to assemble a T-DNA binary vector, based on the crop species they are working with and whether they aim for transient or stable plant transformation.
Schematic showing the two-step process of the ENABLE kit. Step 1 shows insertion of annealed oligos (sgRNA target sites) into pGMF1 and pGMF2 using Golden Gate cloning. pGMF1 and 2 contain a Pol III promoter, sgRNA scaffold, and PolyT stop signal. The sgRNA target sites are inserted between the promoter and scaffold, using compatible BsaI sites (GTTG/ATTG on the five prime of the target site, AAAC on the three prime of target site). Step 2 shows the creation of a complete plasmid using the pGMF3 backbone and Golden Gate cloning. This creates a plasmid with a selection marker (from pGMF5 or pGMF6), the Cas9 cassette (from pGMF4), sgRNA cassette 1 (From pGMF1), and sgRNA cassette 2 (from pGMF2). Compatible restriction sites (using BbsI) from left to right are TGCC, GCAA, ACTA, TTAC, and GGGA.
Figure 2: A two-step, Golden Gate-based cloning method enables easy assembly of a binary T-DNA plasmid containing a selection marker for stable or transient transformation, a Cas9 gene, and two sgRNA genes.

Kit Documentation

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:

"The ENABLE® Gene Editing in planta toolkit was a gift from Kate Creasey Krainer, Vladimir Nekrasov, and Nicola Patron (Addgene kit #1000000270)."

For your Reference section:

Developing a Molecular Toolkit to ENABLE all to apply CRISPR/Cas9-based Gene Editing in planta. Abate BA, Hahn F, Chirivì D, Betti C, Fornara F, Molloy JC, Creasey Krainer KM. bioRxiv. 2025 November 13. doi: 10.1101/2025.11.09.687425. Article (Link opens in a new window)

The ENABLE® Gene Editing in planta Toolkit contains eight plasmids. Please refer to the individual plasmid pages below for more details on each plasmid in this kit:

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