KANEKA CORPORATION

KANEKA New breeding technology New Business Development Division
Main products
Gene delivery Technology Target crops Main products (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
Transformation by Agrobacterium
(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)		 Outline Know-hows and
services provided
iPB Wheat,Barley
Rye, corn
Soy,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
(validated)		 Outline Know-hows and
services provided		 Technologies recommended
for use with
PureIntro™ Rice Highly efficient transformation and genome editing of Japonica and Indica varieties
  • Specialized protocols: optimized experimental conditions and know-how packages
  • Hands-on technical training
  • Super-Ternary Vector Technology: Improved transformation efficiency
PureUpgrade™ Maize Enabling transformation and genome editing in recalcitrant commercial varieties and inbred lines
  • Super-Ternary Vector Technology: Improved transformation efficiency
  • WOX5b Technology: Improved regeneration efficiency
PureWheat™ Wheat Enabling transformation and genome editing in commercial varieties worldwide
  • Super-Ternary Vector Technology: Improved transformation efficiency
  • CB1 Technology: Improved regeneration efficiency
PureBarley™ Barley Highly efficient transformation and genome editing
  • Specialized protocols: optimized experimental conditions and know-how packages
  • Super-Ternary Vector Technology: Improved transformation efficiency
PureSorghum™ Sorghum Enabling transformation and genome editing in recalcitrant varieties
  • Super-Ternary Vector Technology: Improved transformation efficiency

Morphogenic regulators for efficient and genotype independent transformation

Technology Outline Know-hows and
services provided		 Technologies recommended
for use with
CB1
  • Improve regeneration efficiency in Agrobacterium-mediated transformation and genome editing
  • Able to obtain fertile plants even when its expression unit was integrated into the plant genome
  • Target crop: (Proven) Wheat, Maize, Rye. (Under demonstration) Rice
  • CB1 gene cloned in plasmid vector and its sequence information
  • Protocol
  • Pure Technologies: Efficient Agrobacterium-mediated transformation
  • Super-Ternary Vector Technology: Improved transformation efficiency
WOX5b
  • Improve regeneration efficiency in Agrobacterium-mediated transformation and genome editing
  • Able to obtain fertile plants even when its expression unit was integrated into the plant genome
  • More maize optimized and less deleterious than CB1 technology
  • Target crop: (Proven) Maize
  • WOX5b gene cloned in plasmid vector and its sequence information
  • Protocol

Vector technologies

Technology Outline Know-hows and
services provided		 Technologies recommended
for use with
pLC41 A binary vector with excellent stability for transferring long DNA strands
  • Plasmid vector and its sequence information
  • Protocol
 Can be combined with any Agrobacterium-mediated transformation method in principle.
pVGW9 A plasmid which promotes Agrobacterium's infection ability
  • 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.
Example : NLRseek project

Patents and publications (iPB Technology)

Publications

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 (accepted) 2024 DNA-free and genotype-independent CRISPR/Cas9 system in soybean

Patents and publications (tissue culture/transformation technologies)

Publications

Article Title
Plant J, 6:271-82, 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA
Nat Biotechnol, 14:745-50, 1996. High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens
Plant J, 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 Mol Biol, 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 Physiol, 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
Front Plant Sci, 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 Mol Biol, vol 1223, p.155-67 & p. 189-98, 2015. Wheat (Triticum aestivum L.) Transformation Using Immature Embryos
Theor Appl Genet, 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