Transgenic groundnut for drought tolerance

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1. Freedom to Operate (FTO) A FTO study was conducted for Indian & US jurisdictions on various components including DREB1A, NOS Promoter, npt II Marker, rd29A Promoter, NOS Terminator. It was observed from the preliminary FTO study that there is no granted Indian patent on these components exists till date and hence the research activity is not infringing any patent rights in India. IMODInclusive Market Oriented Development Innovate Grow Prosper Timelines and Cost R&D Event Selection BRL 1 BRL 2 Introgression In-House PTTC & Partners Current Stage Event Approval 500,000438,667 410,000 206,000205,556 2013/14 2014/152011/12 Translational Research FTO analysis conducted for Indian and US jurisdictions on various technology components indicated non-infringement of any patent rights Product Development Cycle A A B Homology to Known allergens, toxins, anti-nutrients Blast Searches Heat Stability Acute Oral Toxicity in Rats/Mice (14 Days) Pepsin Digestibility Inheritance Over Multiple Generations RT-PCR, Southern, qPCR data available as above Western & ELISA to be performed with Ab (A) produced from the protein Pedigree Map Documentation from the data recorded from T1 to T5 Sequence data of inserted genetic material & of the flanking regions Identification of ORFs Genome Walking Organisation of inserted genetic material PCR & Sequencing Net House Evaluation Field trials Season 1 Field trials Season 2 Exp. Level of Protein ELISA Reproductive & Survival Biology Phenotyping Site 1 Site 2 Site 3 Site 1 Site 2 Site 3 Composition al Analysis Impact on Non-Target Org. Tier 1 Impact on Non-Target Org Tier 2 Livestock Feeding Studies Number of Insertion Sites qPCR data available as above Antibody Production Protein Equivalence Study Functional, Structural and Biochemical Equivalence Plant Protein purification Purification & scale up of protein from Bacterial / other systems PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR Transformation T0 T1 T2 T3 T5T4 B Construct preparation R&DEventSelectionBRL-I BRL- II Sub-Chronic Feeding Study in Rodents (90 Days) Molecular Studies Field Studies Protein Studies Field trials Gene Flow Aggressiveness & Weediness LOD 0.01% Strictly Confidential A A B Homology to Known allergens, toxins, anti-nutrients Blast Searches Heat Stability Acute Oral Toxicity in Rats/Mice (14 Days) Pepsin Digestibility Inheritance Over Multiple Generations RT-PCR, Southern, qPCR data available as above Western & ELISA to be performed with Ab (A) produced from the protein Pedigree Map Documentation from the data recorded from T1 to T5 Sequence data of inserted genetic material & of the flanking regions Identification of ORFs Genome Walking Organisation of inserted genetic material PCR & Sequencing Net House Evaluation Field trials Season 1 Field trials Season 2 Exp. Level of Protein ELISA Reproductive & Survival Biology Phenotyping Site 1 Site 2 Site 3 Site 1 Site 2 Site 3 Site 1 Site 2 Site 3 Site 1 Site 2 Site 3 Composition al Analysis Impact on Non-Target Org. Tier 1 Impact on Non-Target Org Tier 2 Livestock Feeding Studies Number of Insertion Sites qPCR data available as above Antibody Production Protein Equivalence Study Functional, Structural and Biochemical Equivalence Plant Protein purification Purification & scale up of protein from Bacterial / other systems PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR PCR, Southern, qPCR, RT-PCR Transformation T0 T1 T2 T3 T5T4 B Construct preparation R&DEventSelectionBRL-I BRL- II Sub-Chronic Feeding Study in Rodents (90 Days) Molecular Studies Field Studies Protein Studies Field trials Gene Flow Aggressiveness & Weediness LOD 0.01% Strictly Confidential Transgenic Groundnut for Drought Tolerance Roadmap for Product Development A model depicting product development and dissemination following the regulatory approval for the selected event(s) PTTC ICRISAT NARS / SAUs KVKs / ARS State Seed Corporation Farmers Non-Market Market Technology Provider Product Development & regulatory approvals Varietal Development & Seed Production Product Dissemination PTTC ICRISAT NARS / SAUs KVKs / ARS State Seed Corporation Farmers Non-Market Market PTTC ICRISAT NARS / SAUs KVKs / ARS State Seed Corporation Farmers Non-Market Market Technology Provider Product Development & regulatory approvals Varietal Development & Seed Production Product Dissemination Preliminary Biosafety Data Bioinformatics studies for potential allergenicity indicated absence of any homology of recombinant protein with known toxin / allergen Overview Drought SignalTransduction Transcription/Translation PromoterAPromoterA PromoterBPromoterB PromoterCPromoterC PromoterDPromoterD SignalPerceptionSignalPerception GeneAGeneA GeneBGeneB GeneCGeneC GeneDGeneD DRE PlantCell GeneExpression Stressresponse&tolerance RegulatoryProtein (TranscriptionalActivator) DREB1ADREB1Ard29A-Prd29A-P Drought SignalTransduction Transcription/Translation PromoterAPromoterA PromoterBPromoterB PromoterCPromoterC PromoterDPromoterD SignalPerceptionSignalPerception PromoterAPromoterA PromoterBPromoterB PromoterCPromoterC PromoterDPromoterD SignalPerceptionSignalPerception GeneAGeneA GeneBGeneB GeneCGeneC GeneDGeneD GeneAGeneA GeneBGeneB GeneCGeneC GeneDGeneD DRE PlantCell GeneExpression Stressresponse&tolerance RegulatoryProtein (TranscriptionalActivator) DREB1ADREB1Ard29A-Prd29A-P Improved drought tolerance Improved seed and pod yield (upto 26%) Higher root dry weight and root length density at deeper soil (40-90cm) Improved drought tolerance indices (higher relative harvest index (RHI) and yield reduction under drought stress) Sep 2011

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