Freedom to Operate (FTO) Timelines and Cost R&D Event Selecon BRL 1 BRL 2 Introgression In-House PTTC & Partners Current Stage Event Approval 500,000 438,667 410,000 206,000 205,556 2013/14 2014/15 2011/12 Translaonal 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 T 1 to T 5 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 T 0 T 1 T 2 T 3 T 5 T 4 B Construct preparation R&D Event Selection BRL-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% 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 T 1 to T 5 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 T 0 T 1 T 2 T 3 T 5 T 4 B Construct preparation R&D Event Selection BRL-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% 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 PTTC 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 Signal Transducon Transcripon / Translaon Promoter A Promoter A Promoter B Promoter B Promoter C Promoter C Promoter D Promoter D Signal Percepon Signal Percepon Gene A Gene A Gene B Gene B Gene C Gene C Gene D Gene D DRE Plant Cell Gene Expression Stress response & tolerance Regulatory Protein (Transcriponal Acvator) DREB1A DREB1A rd29A-P rd29A-P Drought Signal Transducon Transcripon / Translaon Promoter A Promoter A Promoter B Promoter B Promoter C Promoter C Promoter D Promoter D Signal Percepon Signal Percepon Promoter A Promoter A Promoter B Promoter B Promoter C Promoter C Promoter D Promoter D Signal Percepon Signal Percepon Gene A Gene A Gene B Gene B Gene C Gene C Gene D Gene D Gene A Gene A Gene B Gene B Gene C Gene C Gene D Gene D DRE Plant Cell Gene Expression Stress response & tolerance Regulatory Protein (Transcriponal Acvator) DREB1A DREB1A rd29A-P rd29A-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