Producing Transgenic Plants - ris.org.in · Brinjal 4 5 9 Black Pepper 3 0 3 ... 愀渀搀...
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DBT P l a t f o r m f o r T r a n s l a t i o n a l R e s e a r c h o n T r a n s g e n i c C r o p s ( P T T C ) Fifth Asian Biotechnology and Development Conference, Sri Lanka Dec. 15-18, 2010 International Crops Research Institute for the Semi-Arid Tropics Patancheru 502 324, Andhra Pradesh, India K i r a n K . S h a r m a G e n e t i c T r a n s f o r m a t i o n L a b o r a t o r y , I C R I S A T k . s h a r m a @ c g i a r . o r g P o o j a B h a t n a g a r - M a t h u r , I C R I S A T S . R . R a o - D e p a r t m e n t o f B i o t e c h n o l o g y . G o I K i r a n K . S h a r m a G e n e t i c T r a n s f o r m a t i o n L a b o r a t o r y , I C R I S A T k . s h a r m a @ c g i a r . o r g P o o j a B h a t n a g a r - M a t h u r , I C R I S A T S . R . R a o - D e p a r t m e n t o f B i o t e c h n o l o g y . G o I
Transcript of Producing Transgenic Plants - ris.org.in · Brinjal 4 5 9 Black Pepper 3 0 3 ... 愀渀搀...
DBT
Platform for Translational Research on Transgenic Crops (PTTC)
Fifth Asian Biotechnology and Development Conference, Sri Lanka Dec. 15-18, 2010
International Crops Research Institute for the Semi-Arid Tropics
Patancheru 502 324, Andhra Pradesh, India
Kiran K. SharmaGenetic Transformation Laboratory, ICRISAT
Pooja Bhatnagar-Mathur, ICRISATS.R. Rao- Department of Biotechnology. GoI
Kiran K. SharmaGenetic Transformation Laboratory, ICRISAT
Pooja Bhatnagar-Mathur, ICRISATS.R. Rao- Department of Biotechnology. GoI
Producing Transgenic Plants•
Efficient tissue culture system for regenerating shoots•
Introduction of gene into plant cells•
Selection of transformed cells or tissues•
Regeneration of putatively transformed whole plants
•
Transfer to greenhouse and advancement of generations•
Molecular and genetic characterization•
Selection of events under greenhouse conditions
•
Contained field testing under natural conditions•
Open field testing for agronomic performance•
Food and Environmental safety•
Release, introgression into new varieties, & Commercialization
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DBT
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Indian Research Scenario
The figures include basic research
Crops Pub. Pvt. TotalFIELD CROPSWheat 1 0 1Rice 30 5 35Ragi 2 0 2Sorghum 10 2 12Mustard 5 1 6Maize 2 0 2
Subtotal 50 8 58VEGETABLESBrinjal 4 5 9Black Pepper 3 0 3Bell Pepper 1 0 1Okra 0 6 6Cabbage 2 2 4Carrot 1 0 1Cauliflower 2 3 5Chillies 4 0 4Tomato 20 7 27Potato 8 0 8
Subtotal 45 23 68
Crops Pub. Pvt. TotalFRUITSBanana 3 0 3Citrus 1 0 1Muskmelon 1 0 1Pomegranate 1 0 1Watermelon 2 0 2
Subtotal 8 0 8OTHER CROPSChickpea 5 2 7Pigeonpea 13 1 14Groundnut 9 0 9Cassava 2 0 2Soybean 2 1 3Sunflower 1 1 2Mulberry 0 0 0Cotton 11 50 61Coffee 1 0 1Cardamom 1 0 1
Sutotal 45 55 99Total 148 86 234
Source: IGMORIS and public domain
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Transgenic Crops Research in India
The figures include basic research Source: IGMORIS and public domain
• 6 technologies (cotton) already approved (5 private & 1 public)
• 4 technologies waiting for commercial approval (all private)
• 29 technologies approved for field testing in 2010 (4 in BRL stages and all belong to private)
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AgBiotech driven by lengthy Product Development Cycle and Large Investment Process*
Year 0 1 2 4 6 8 93 5 7 10DiscoveryDiscovery Phase IPhase I Phase IIPhase II Phase IIIPhase III Phase IVPhase IV
Gene/Trait IdentificationGene/Trait Identification Proof of ConceptProof of Concept Early DevelopmentEarly Development Advanced DevelopmentAdvanced Development Regulatory SubmissionRegulatory Submission
• High throughput screening
• Model crop testing
• High throughput screening
• Model crop testing
• Gene optimization• Crop
transformation
• Gene optimization• Crop
transformation
• Traitdevelopment
• Pre-regulatory data
• Large scale transformation
• Trait development
• Pre-regulatory data
• Large scale transformation
• Trait integration• Field testing• Regulatory data
generation
• Trait integration• Field testing• Regulatory data
generation
• Regulatory submission
• Seed bulk-up• Pre-marketing
• Regulatory submission
• Seed bulk-up• Pre-marketing
• Regulatory submission
• Seed bulk-up• Pre-marketing
$2-5M(5%)
$5-10M(25%)
$10-15M(50%)
$15-30M(75%)
$20-40M(90%)On Average:
Time to market: 8-10 yearsTotal expense: ~$100M
On Average:On Average:Time to market: 8-10 yearsTotal expense: ~$100M
On Average:Time to market: 8-10 yearsTotal expense: ~$100M
Spen
ding
(Pro
babi
lity
of S
ucce
ss in
%)
* Numbers (time duration, spending, and probability of success) are all estimates. The actual for individual projects vary. Source: S.R. Rao-DBT
Presenter�
Presentation Notes�
We invest tremendous efforts in biotech R&D. From a macro view, here you see the time to market and related investment required to develop just one potential product in a pipeline. It takes an average of 8-10 years for a product to reach commercialization and a potential of $100 million dollars in expense. From discovery to launch, we have four phases to represent different stage of R&D process, you can see we use phase concept to describe our pipeline status in future slides. The further along a product progresses through the R&D pipeline, the more likely its success. �
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Distribution of Total R&D Expenditure in Science and Technology
PercentagePublic Private
India 95 05Mexico 88 12Indonesia 96 04Zimbabwe 86 14USA 30 70Switzerland 26 74
Source: S.R. Rao-DBT
Developing new Biotechnology is not cheap!Most current research is being done by private
sector companies in the developed world
• In most of the developing countries, public sector invests in research and development.
• Private sector investment is beginning to increase due to globalization and reforms in some of them.
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• Failure to test the concepts on a large scale
• Failure to translate concepts into commercial products
• Barriers of IP/Trade/interest of private sector
• Biosafety regulations/Food safety
• Risk assessment and risk management
• Training & Capacity building
• Partnerships for product development & deployment
• Commercialization limitations, a serious barrier
Bottlenecks
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Problems in commercialization
Lack of proper understanding of issues related to:
Intellectual property rights
Regulatory aspects
Roadmap for lab to farmers’ field
Market demands
Public-privatepartnerships in R&D
Priority setting for Transgenic Plants
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1. Overcome delays in moving GM crops to farmers.
2. Food safety & Research issues linked to risk assessment and risk management.
3. Training on the development and deployment aspects.
4. Public awareness (media workshops; training of policy makers, trainers, NGOs etc…. ).
5. Socioeconomics & potential impact.
6. Partnerships for product development & deployment [Commercialization]
i. Public sector partners (groundnut and chickpea)ii. Transgenic Research Consortia (hybrid pigeonpea; other crops)iii. Facilitate technology development & commercialization (ASP@ICRISAT))iv. Facilities for translation of transgenic products (PTTC)
SolutionsSolutionsDeployment of transgenicsDeployment of transgenics
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Technology TransferTechnology Transfer
Develop strong relationships between:Basic Research Technology Development
Technology Transfer Technology Diffusion
Focus on human resource development.
Create congenial environment for encouraging R & D through the development of infrastructure and incentives.
Interface public and private sectors for enhanced synergies in technology development and deployment.
Presenter�
Presentation Notes�
-R&D in the public sector has tended not to interface with private enterprise and so is disconnected from the broader market developments and demands. Underlying assumption that research, technology development, technology transfer, and technology use were a sequential tasks, has limited the emergence of more iterative relationships between science, the market, and technology users. WTO means that Indian agricultural and the agro-based industries must become more competitive, both in terms of price, but also in terms of quality. In many sectors a quality reputation is yet to be built. Indian wheat prices for example are significantly higher than international prices due to policy distortions in India. Entry into global markets and international competition requires rapid technical change in both production and post-harvest arenas. Existing institutional arrangements make this difficult, particularly so where scientific expertise cuts across exiting organizational boundaries and where partnership between public and private sectors are required. New, flexible and evolving multi-agency groupings are required. Environmental degradation and loss of biodiversity are emerging as major threats to the sustainability of current agricultural development strategies. Science needs to provide new strategic opportunities that go beyond ameliorating existing constrains, and contribute to an new development paradigm that will prove sustainable in the long run. Rural poverty remains, with more than 450 million people surviving on less than a dollar a day. Agricultural development must address this problem through technological and institutional changes, that draw on the creativity of science, the public and private sectors in ways that genuinely enfranchise the poor. �
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Principles1. Create and charter an entity with express purpose of
“translating” genetic engineering research into a practical, value adding technology
2.The entity would embody the requisite scientific andbusiness skills that are appropriately balanced
MissionTo “translate transgenic technology and harness its products to meet the needs
of agricultural growth”
An ICRISAT-DBT InitiativePTTC
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To promote an existing transgenic technology into a value added product
of commercial use
Translational Research?
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The main drivers of innovation are priorityand potential new technology
PTTCinterventionpoint
Implementation
R & D of a new technology
Assessment of the technology
Proof of concept
Assess market potential
Design strategy
Innovation= Invention + Commercialization
Roadmap
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Concept
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Partnership
(Public-Private /Public-Public)
Partnership
(Public-Private /Public-Public)
Partnership
(Public-Private /Public-Public)
PTTC• Research & Development• Field Evaluation• Safety Assessments• IP Evaluation• Regulatory Approvals
PTTC• Research & DevelopmentResearch & DevelopmentResearch & DevelopmentResearch & DevelopmentResearch & DevelopmentResearch & DevelopmentResearch & DevelopmentResearch & Development• Field EvaluationField EvaluationField EvaluationField EvaluationField EvaluationField EvaluationField Evaluation• Safety AssessmentsSafety AssessmentsSafety AssessmentsSafety AssessmentsSafety AssessmentsSafety AssessmentsSafety AssessmentsSafety AssessmentsSafety Assessments• IP EvaluationIP EvaluationIP EvaluationIP EvaluationIP EvaluationIP EvaluationIP Evaluation• Regulatory ApprovalsRegulatory ApprovalsRegulatory ApprovalsRegulatory ApprovalsRegulatory ApprovalsRegulatory ApprovalsRegulatory Approvals
PTTC• Research & Development• Field Evaluation• Safety Assessments• IP Evaluation• Regulatory Approvals
Selected Technology from Public Sector Research Institute
Selected Technology from Public Sector Research Institute
Selected Technology from Public Sector Research Institute
FundingAgencies FundingAgencies Funding Agencies
ProductProductProduct
Partners(ICAR, ICMR,
CSIR, SAUs, etc.)
Partners(ICAR, ICMR,
CSIR, SAUs, etc.)
Partners(ICAR, ICMR,
CSIR, SAUs, etc.)
Regulatory Bodies
Regulatory Bodies
Regulatory Bodies
International Experts
International Experts
International Experts
ServicesServicesServices
Operational Model
PTTCSustenance
Fund
PTTCSustenance
Fund
PTTCSustenance
Fund
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To develop and deploy state-of-the-art infrastructure for conduct of transgenic research.
To act as a clearinghouse for technology inputs,transgenic research leads/ prototypes with proof of concept derived from Indian research institutes,universities, and other likely sources.
To evaluate specific concepts, ideas and technologies, and advance the promising transgenic events through a development cycle with adequate safety assessments.
To “evolve” the technology to a point where a practical application can be demonstrated, and transfer this “evolved” technology for product development and distribution to appropriate agencies
Objectives
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PTTC Executive Committee
Technology Advancement Advisory Committee (TAAC) - Representatives from ICRISAT, DBT, ICAR, SAUs, Private Sector, Investors, NGOs, etc.
International Advisory Board – Eminent Scientists, Policy makers, Venture capitalists, Technology Managers, IP Experts, etc.
Organization
Dr. William DarDirector General
ICRISAT
Dr. Kiran K.SharmaDirector, PTTC & Principal
Scientist. ICRISAT
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Infrastructure & Facilities
• State-of-the-art infrastructure for transgenic crop development on an area of 50,000 Sq. Ft
• High-throughput transformation facility
• Well equipped molecular biology laboratory
• Analytical laboratory & Instrumentation facility
• Plant pathology & virology laboratory
• Insect rearing facility
• P2 level contained and regular greenhouses
• Contained Fields
P2 level contained and regular greenhouses
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• Genetic transformation
• Genotyping: PCR, Southern analysis, qPCR with liquid handling systems, proteomics, etc.
• Phenotyping: Screening for resistance to aflatoxin, virus, foliar diseases; tolerance to drought & salinity; etc.
• Food & Feed Safety Assessment: heat stability, pepsin digestibility, homology search, etc.
• Environmental Safety Assessment: Gene flow studies, aggressiveness & weediness Potential, effect on non target organisms (tier 2), etc
• Field Trials: Confined Field Trials (BRL I & BRL II)
• IP Management: FTO Analysis,
• Product Dossier Preparation: Crop biology document, Technology dossier, Biosafety dossier
Research Capacities
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Independent evaluation of trait-specific transgenic events
Examine IPR issues in transgenic product development
Develop product dossiers for commercialization
Create specific projects with defined milestones and end-points
Coordinate and conduct the evaluation of transgenic events for biosafety studies
Introgression of the desirable transgenic events
Identify partnerships for sharing mechanisms for marketing of the final “product”
Activities
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Roadmap
Need & benefits identified by SAC/TAC at PTTC
Potential technology identified by PTTC
EOI/Application received
Complete review and due diligence carried out
Resource needs and sources to be identified and sought
Establishment of the process (Event selection/risk assessment etc.)
Adoption and implementation of the process towards commercialization
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FTO Analysis
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Product Development Pipeline
A
A
B
Homology to Known allergens, toxins,
anti-nutrients Blast Searches
HeatStability
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 BConstruct
preparation
R&
DE
ven
t Sele
ction
BR
L-I
BR
L-
II
Sub-Chronic Feeding Study in Rodents
(90 Days)
Molecular Studies Field Studies
Protein StudiesField trials
Gene Flow
Aggressiveness & Weediness
LOD 0.01%
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• Project Partners –ICRISAT & DBT
• Activity Partners -MoA with NARS like NIN, ITRI, SAUs, etc. and private sector organisations on need basis
• Preliminary discussion with some org. already completed
• MoU with Associates/ Consultants arebeing executed
PTTC Network
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Product commercialization in a competitive mode
Public Funding for Public Private Partnership (SBIRI,BIPP)
Proposal to be developed jointly with Applicant in a
project mode
Public Funding (DBT, ICAR)
Private Industry (Consortia etc.)
Potential technology identified for product development
Contract Research Services
Client’s own Funding
Proposal to be developed jointly with Applicant in a
project mode
Funding Mechanism
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PTTC Services
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Research Service Packages
Genomics• Stable integration over multiple
generations• copy Number and number of insertion
sites• Organization of Inserts• Flanking regions study• Bioinformatics analysis of novel ORFs
putative chimeric proteins• T-DNA sequencing• Absence of plasmid backbone
Proteomics• Protein Characterization & Scale-up• Homology Studies• Heat Stability• Pepsin Digestibility
Environmental Assessment Studies
• Plant pest potential• Out crossing• Gene Flow Studies• Aggressiveness and weediness• Impact on NTOs (Tier-1 & Tier 2)• LOD 0.01%
Toxicity and Allergenicity• Acute oral toxicity test in rodents• Sub chronic feeding test in rodents• Primary skin irritation test in rabbits• Mucous membrane irritation study in
female rabbits
Compositional AnalysisAsh, Total fat, moisture, Protein, Crude fibre, Acid detergent fibre, amino acid profile, Total lipids, Fatty acid profile, Vitamin profile, Mineral profile
Livestock Feeding StudiesRepresentative Animals – Goat, Chicken, Cow / Buffalo, Fish, etc.
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Referral Documents
Forms
• Expression of Interest – Public & Private Sector
• Evaluation of Technology Roadmapping
Agreements
• Memorandum of Understanding
• Memorandum of Agreement
Service Agreement
Partnership Agreement
• Non-Disclosure Agreement
• Material Transfer Agreement
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• Flyers, FAQs, Posters, Website, etc.
• Stakeholder meetings, Media Workshops, Biosafety workshops etc.
• Crop biology documents
Outreach
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Outcomes
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Looking aheadLooking aheadLooking ahead
PTTC has the potential to evolve into a “leading edge” technology translational facility
It could serve as a global model for the utilization of transgenictechnologies and their products (North-South & South-South)
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Thank you
