KANEKA CORPORATION

Solutions (including licensing and technical services)

Gene delivery Technology	Target crops	Solutions (services)
iPB technology (in planta particle bombardment)	Wheat, Barley, Rye, Soybean, etc.	1.Technology licensing 2.Contract transformation service 3.Large scale gene screening 4.Joint development of traits and varieties
Agrobacterium-mediated transformation (inherited from JT and developed further by KANEKA)	Wheat, Rice, Maize, Potato, Soybean, etc.

1.Technology licensing

In Planta Particle Bombardment (iPB) Technology

Technology	Crop species (validated)	Overview	Items and services
iPB	Wheat, Barley, Rye, Maize, Soybean and Melon	Highly efficient genome editing and gene transfer without the need for tissue culture	Specialized protocols: optimized experimental conditions and know-how packages Hands-on technical training

Agrobacterium mediated transformation technologies

Technology	Crop species	Overview	Items and services	Technologies recommended for use with
PureUpgrade Solution	Maize	An integrated maize transformation solution composed of: PureUpgrade technology (core transformation system) WOX5b technology (proprietary regeneration factor optimized for maize) Super Ternary Vector technology Enabling efficient gene delivery and trait development in recalcitrant maize varieties	Specialized protocols: optimized experimental conditions and know-how packages Hands-on technical training	All recommended technologies are included as part of the solution
PureWheatPlus Solution	Wheat	An enhanced wheat transformation solution composed of: PureWheatPlus technology (based on our improved transformation system) CB1 technology (proprietary regeneration factor) Enabling efficient gene introduction and trait development in diverse wheat varieties		pLC41: A highly stable binary vector suitable for the transfer of large or structurally complex DNA
PurePotato	Potato	Enabling highly efficient transformation of a wide range of commercial varieties and subsequent genome editing		Super-Ternary Vector Technology: Improved transformation efficiency
PureSoy	Soybean	Reduces genotype dependency, enabling transformation and genome editing in diverse commercial varieties.
PureIntro	Rice	Highly efficient transformation supporting genome editing across Japonica and Indica rice
PureBarley	Barley	Highly efficient transformation, enabling genome editing applications.	Specialized protocols: optimized experimental conditions and know-how packages
PureSorghum	Sorghum	Enabling transformation and genome editing in recalcitrant varieties	Specialized protocols: optimized experimental conditions and know-how packages

Morphogenic regulators for efficient and genotype independent transformation

Technology	Overview	Items and services	Technologies recommended for use with
CB1	Improves regeneration efficiency in Agrobacterium-mediated transformation and genome editing. Enables the generation of fertile plants even when its expression cassette is integrated into the plant genome. Target crops: 　Proven: Wheat, maize, rye 　Under demonstration: Rice	CB1 gene cloned in plasmid vector and its sequence information Protocol	Pure Technologies: Technologies optimized for each crop species for efficient Agrobacterium-mediated transformation. Super-Ternary Vector Technology: Enhances Agrobacterium-mediated T-DNA transfer and improves binary vector stability.
WOX5b	Improves regeneration efficiency in Agrobacterium-mediated transformation and genome editing. Enables the generation of fertile plants even when the expression cassette is integrated into the plant genome. Optimized for maize with reduced deleterious effects compared to CB1. Target crop: 　Proven: Maize	WOX5b gene cloned in plasmid vector and its sequence information Protocol

Vector technologies

Technology	Overview	Items and services	Technologies recommended for use with
pLC41	A highly stable binary vector enabling efficient transfer of long or structurally complex DNA.	Plasmid vector and its sequence information Protocol	Can be combined with any Agrobacterium-mediated transformation method in principle.
pVGW9	Enhances Agrobacterium-mediated T-DNA transfer into the plant genome.	Plasmid vector and its sequence information Protocol
Super-Ternary Vector System	Combination of pLC41 and pVGW9	Plasmid vectors and their sequence information Protocol

2.Contract transformation service

Crop species	Outline	Note
Wheat, Maize, Rice,Soybean	Contracted transformation with our proprietary Phenotypic and molecular biological analyses are also available Deliver transformant seeds with target genes to customers	Customer varieties can be used if they are allowed to be imported into Japan.

Possible arrangement for 2: contract transformation

3.Large scale gene screening

Crop species	Outline	Note
Monocots	Highly efficient transformation enables screening a large number of transgenic events.	Customer varieties can be used if they are allowed to be imported into Japan.

This work was published in Nature Plants in 2025.

publications _{(iPB Technology)}

Article	Title
Scientific Reports 7: 11443, 2017.	An in planta biolistic method for stable wheat transformation
Scientific Reports 8: 14422, 2018.	Biolistic-delivery-based transient CRISPR/Cas9 expression enables in planta genome editing in wheat
Plant Biotechnology 37(2): 171-176, 2020.	In planta particle bombardment (iPB): A new method for plant transformation and genome editing
Frontiers Plant Science 2021.648841, 2021.	In planta Genome Editing in Commercial Wheat Varieties
Plant Physiology 188(4) 1838-1842, 2022	Introduction of a second “Green Revolution” mutation into wheat via in planta CRISPR/Cas9 delivery
Plant Biotechnology Journal 21: 668-670, 2023	Precise in planta genome editing via homology-directed repair in wheat
Plant Physiology 196(4): 2320–2329, 2024	DNA-free and genotype-independent CRISPR/Cas9 system in soybean
Plant Physiology 196(4): 2263–2265, 2024	Killing two birds with one stone: A breakthrough in transgene-free gene editing in soybean
Frontiers in Genome Editing, 18 June 2025	A long shelf-life melon created via CRISPR/Cas9 RNP-based in planta genome editing

publications _{(Plant tissue culture and Agrobacterium mediated transformation)}

Article	Title
The Plant Journal 6:271-82, 1994.	Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA
Nature Biotechnology 14:745-50, 1996.	High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens
The Plant Journal 6:271-82, 1994.	Vectors carrying two separate T-DNAs for co-transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers
Plant Molecular Biology 35:205-18, 1997.	Transformation of rice mediated by Agrobacterium tumefaciens
Molecular Breeding 14:309-20, 2004.	Suppression of transfer of non-T-DNA ‘vector backbone’ sequences by multiple left border repeats in vectors for transformation of higher plants mediated by Agrobacterium tumefaciens
Plant Cell Tissue Organ Culture 85:271-83, 2006.	Improved protocols for transformation of indica rice mediated by Agrobacterium tumefaciens
Plant Cell Tissue Organ Culture 87(3):233-43, 2006.	Improved frequency of transformation in rice and maize by treatment of immature embryos with centrifugation and heat prior to infection with Agrobacterium tumefaciens
Nature Protocols 2(7):1614-21, 2007.	Agrobacterium-mediated transformation of maize
Plant Physiology 145:1155-60, 2007.	Current Status of Binary Vectors and Superbinary Vectors
Nature Protocols 3(5):824-34, 2008.	Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed
Historical Technology Developments in Plant Transformation. Bentham Science p. 55-76, 2011.	Transformation Vectors and Expression of Foreign Genes in Higher Plants
Frontiers in Plant Science 5:628, 2014.	Progress of cereal transformation technology mediated by Agrobacterium tumefaciens
Advances in Wheat Genetics: From Genome to Field. New York: Springer p. 167-73, 2015.	High Efficiency Wheat Transformation Mediated by Agrobacterium tumefaciens
Agrobacterium Protocols, 3rd edition Vol 1, Methods in Molecular Biology, vol 1223, p.155-67 & p. 189-98, 2015.	Wheat (Triticum aestivum L.) Transformation Using Immature Embryos
Theoretical and Applied Genetics 133:1291-1301, 2020.	High-throughput phenotypic screening of random genomic fragments in transgenic rice identified novel drought tolerance genes
Plant Biotechnology 37:121-28, 2020.	Tissue culture protocols for gene transfer and editing in maize (Zea mays L.)
Nature Plants 8: 110-117, 2022.	The gene TaWOX5 overcomes genotype dependency in wheat genetic transformation
Nature Plants 11:2100-2114, 2025.	Discovery of functional NLRs using expression level, high-throughput transformation and large-scale phenotyping