Working With Recombinant DNA

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Working With Recombinant DNA

• 2007 Information Session
• Professor Marilyn Anderson
• Chair, Genetic Manipulation Supervisory Committee

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What is a GMO?

• A genetically modified organism
• An organism that has inherited traits from a GMO

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Organisms that are not GMOs

• Organisms declared by the Regulator not to be
  GMOs
• Humans that have undergone somatic cell gene
  therapy
• Mutant organism in which mutational event did not
  involve the introduction of foreign nucleic acid
• Protoplast fusion between non-pathogenic bacteria
  or yeast

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Organisms that are not GMOs

• Recombinant organism formed by integration of DNA
  sequences of a genetic element that occurs
  naturally in the species and moves sporadically
  between genome sites
• A plant formed by embryo rescue or protoplast
  fusion
• Organism that results from the exchange of DNA if
  it is from the same species or if DNA exchange
  naturally occurs between those species

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How is Gene Technology Research Regulated?

• Gene Technology Act 2000 and Gene Technology
  Regulations 2001
• To protect the health and safety of people, and
  to protect the environment by identifying risks
  posed by, or as a result of gene technology, and
  by managing those risks through regulating
  certain dealings with GMOs

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Office of the Gene Technology Regulator (OGTR)

• Administers legislation and assesses risks
  imposed by GMOS
• Accreditation of organisations
• Certification of facilities
• Regulation of human cloning
• Import and transport of GMOs
• Regulation of GM products

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Office of the Gene Technology Regulator (OGTR)

• Informs and advises other regulatory agencies and
  the public about GMOs and GM products
• Three key advisory groups
• Gene Technology Technical Advisory Committee
  (GTTAC)
• Gene Technology Ethics Committee (GTEC)
• Gene Technology Community Consultative Committee
  (GTCCC)

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Genetic Manipulation Supervisory Committee (GMSC)

• Ensures that La Trobe University complies with
  gene technology legislation
• Composition of the GMSC includes scientists
  (including a microbiologist, a molecular
  biologist and/or geneticist), a biological safety
  officer and a lay person

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Duties of the GMSC

• Assessing research proposals and determining
  classification
• Inspecting PC1 and PC2 facilities
• Reporting annually to the OGTR on research being
  conducted at the University
• Training University staff and students on
  complying with OGTR requirements

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What is a Dealing?

• Conducting experiments with a GMO
• Making, developing, producing or manufacturing a
  GMO
• Breeding or propagating a GMO
• Using a GMO to make something that is not a GMO
• Growing, raising or culturing a GMO
• Importing, sorting, transporting or disposing of
  a GMO

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Types of Dealings

• Exempt dealings
• Notifiable low risk dealings (NLRD)
• Licensed dealings
• Not released to the environment (DNIR)
• Released to the environment (DIR)

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Exempt Dealings

• Lowest perceived risk
• All exempt dealings must be conducted in a PC1
  laboratory (at least)
• Requires approval from the GMSC only
• Exempt dealings include
• Production or use of knockout mice
• Any dealing with a whole animal if DNA/RNA has
  been introduced into somatic cells and cant give
  rise to infectious agents

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Exempt Dealings

• Any dealing involving shot-gun cloning of
  mammalian DNA in a host/vector system listed in
  part 2 of the schedule http//scaleplus.law.gov.au
  /html/pastereg/3/1664/0/PR000600.htm
• Any dealing involving a host/vector system listed
  in part 2 of the schedule and providing no more
  than 10L of culture is produced if the donor DNA
• Is not an oncogene
• Is not from micro-organisms that can cause
  disease, or is fully characterized and will not
  increase the virulence or host range of the vector

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Exempt Dealings

• Does not code for a toxin with LD50lt100µg/kg
• Is not uncharacterized DNA from an organism that
  produces toxins
• The donor DNA includes viral sequences but is
  missing at least one gene essential for viral
  replication

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Host/Vector Systems for Exempt Dealings
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Notifiable Low Risk Dealing

• Any dealing involving whole animals that involves
  genetic modification of the genome of the oocyte
  or zygote or early embryo
• Any genetically modified plant providing it cant
  set seed or produce viable pollen that can escape
• Any host and vector not mentioned in schedule 2
  if the host and the vector is incapable of
  causing disease (in humans, animals, plants or
  fungi)

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Dealings Not involving Intentional Release

• All dealings with GMOs that are not exempt
  dealings, NLRDs or on the GMO register, must be
  licensed by the Regulator AND the GMO is
  physically contained and is not released into the
  environment
• The regulator assesses the application by
  preparing a risk assessment and risk management
  plan
• All license conditions must be adhered to
• All named investigators must sign the license

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Dealings involving Intentional Release

• A dealing with a GMO involves the intentional
  release of a GMO into the environment if the GMO
  is intentionally released into the open
  environment, whether or not it is released with
  provision for limiting the dissemination or
  persistence of the GMO or its genetic material in
  the environment
• Examples include
• grow a GM crop as a field trial
• grow a GM crop commercially
• release a GM fish into a waterway

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Decision Tree for Approvals for Dealings with GMOs
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Transportation of GMOs

• Includes all movements of GMO from a certified
  facility to any location outside the certified
  facility
• Includes all movements from a location specified
  in a licence to another location
• You must include transportation of GMO in your
  dealing application
• Transportation of GMO WASTE follows the same
  conditions

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Conditions of GMO Transportation

• The GMO must be wholly contained in a primary
  sealed container and
• The primary sealed container must be packed in a
  secondary sealed unbreakable container
• The secondary container must be labelled to
  indicate it contains GMOs, and the label must
  contain the name and telephone number of a
  contact person should the package be lost or
  damaged
• Following transportation the primary and
  secondary containers must be decontaminated

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Physical Containment 1 (PC1) Facilities

• All exempt dealings must be performed in a PC1
  facility (at least)
• Certified if the OGTR grant special dispensation
  for an NLRD to be conducted in a PC1 facility
• Certified PC1 facilities inspected annually by
  the GMSC
• Staff and students must be trained in PC1
  practices

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PC1 Procedures

• No eating, drinking, smoking or application of
  cosmetics
• No storage of food or drink
• Lab gowns must be worn and removed before leaving
• Closed footwear must be worn
• Safety glasses to be worn where appropriate to
  protect eyes and face from splashes and other
  hazards
• Keep only the minimum required quantities of
  hazardous substances in the laboratory work area

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PC1 Procedures

• Clearly identify and date cultures. Do not store
  cultures for long periods of time on the bench
• Take care to minimize the production of aerosols
  where work is being carried out on the bench
• Clean up spills immediately and decontaminate the
  area
• Decontaminate work benches at least daily and
  after each task is completed
• Segregate specialized waste at point of discard
  and dispose of according to local regulations

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Physical Containment 2 (PC2) Facilities

• Unless granted special dispensation from the OGTR
  all NLRD work must be performed in a PC2 facility
• Certified by the OGTR
• Inspected annually by the GMSC
• Staff and students must be trained in PC2
  practices and a record of this training must be
  kept

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PC2 Procedures

• Keep doors closed when work is in progress
• No mouth pipetting
• No eating, drinking, smoking or application of
  cosmetics
• No storage of food or drink
• Wear lab gowns and remove before leaving
• Sterilise all biological waste
• Wash hands before leaving
• Dont moisten labels with the tongue

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PC2 Procedures

• Equipment that cant be steam sterilized must be
  chemically disinfected after use
• Decontaminate work benches and surfaces with a
  disinfectant solution after spills and when work
  is completed
• Material carried out of the laboratory to an
  autoclave or elsewhere must be in a closed
  unbreakable container
• Minimize the creation of aerosols-sonicate or
  vortex in a biological safety cabinet
• Clearly identify, date and appropriately store
  cultures

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GMO Storage Policy

• All GMOs must be stored in sealed containers that
  are clearly labeled so that GMOs are easily
  identifiable.
• In order to ensure that GMOs are stored lawfully,
  they must be covered by a current dealing, or an
  application to store GMOs for future research.
• GMOs must be stored inside a PC2 facility OR
• in a cold storage cabinet that is either
  lockable or located in a secure area that
  prevents access to the cabinet from persons not
  responsible for its contents

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State and National Ethical Principles

• The Statement of Ethical Principles for
  Biotechnology in Victoria contains 8 principles
  (http//www.health.vic.gov.au/biotechnology/downlo
  ads/statement1794C0.pdf)
• Respect for persons
• Respect for animals
• Respect for the natural environment
• Respect for the public good
• Benefit and harm
• Justice and equity
• Probity
• Accountability

• The National Framework for the Development of
  Ethical Principles in Gene Technology contains 9
  principles (http//www.ogtr.gov.au/rtf/committee/g
  tecethicalprinc.rtf) .

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Changes in the Wind
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Regulations, Policies and Application Forms

• Office of the Gene Technology Regulator (OGTR)
  www.ogtr.gov.au
• Faculty OHS http//www.latrobe.edu.au/molsci/oh
  s/index.html
• Research and Graduate Studies Office
• www.latrobe.edu.au/rgso/ethics

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Further Information

• Dr Julie Blasioli
• Secretary, Genetic Manipulation Supervisory
  Committee
• Ph 9479 2043
• Email gmsc_at_latrobe.edu.au