Foundation Training in Biological Safety

Module 4Genetically modified

organisms

Why GM?

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Gene of interest (insert)

Vector

Host

Protein of interest

What is GM?(e.g. bacterial expression system)

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Procedures not considered as GM

• Mutagenesis- chemical or radiation

• Cell fusion of prokaryotic species which can exchange genetic material through homologous recombination

• Cell fusion of cells of any eukaryotic species, including production of hybridomas

• Self-cloning, where the resulting organism is unlikely to cause disease or harm to humans, animals or plants

Not GM

The GMO (Contained use) Regulations:key duties

• Assess risks created by work to either human health or the

environment presented by GMMs/GMOs

• Review and record risk assessments

• Notification of Class 2 and 3 GMM activities

• Apply the principles of occupational and environmental safety

• Apply the containment measures specified

• Have in place emergency plans

• Report accidents

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GM Regulations

• Where GMMs or GMOs are cultured, stored, used, transported, destroyed or disposed of.

• Contained using barriers to protect the general population and environment– Physical– Chemical – Biological– Management

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Gene of interest (insert)

Vector

Host

Protein of interest

Example of GM(bacterial expression system)

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Risks

• HOST– How pathogenic?– Virulence and infectivity?– Are toxins produced?– Allergenic factors? – Seriousness of exposure? – Survivability outside lab?

                                                                          

E. coli K12, the prokaryotic wimp

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RISKS

• VECTORS– Consider:– Pathogenicity– Transferability of the vector:– Only want vector to transfer target host, achieved

by modifying how a vector transfers

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RISKS

• INSERTS

– Consider: – gene expression e.g. overexpression– hazards posed by the gene product e.g. activity of

expressed protein, toxic, allergenic or pathogenic effects– Different effects depending on the location of the insert– Alteration of existing pathogenic traits?

THE GM (CONTAINED USE) REGULATIONS

Requires that all GMMs are classified in one of four Classes

Class 1 Class 2 Class 3 Class 4

Increasing hazard to human health or the environment

1 2 3 4

Containment Level

Classification of GMM determined by Containment Level required

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GM animals or plants

• Genetic material altered using a method that does not occur naturally. Not chemical or physical mutagenesis

• Stable integration of the inserted GMM into the genome

• Direct injection of naked DNA to elicit an immune response not GM

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RISK ASSESSMENT APPROACH FOR GM ANIMALS AND PLANTS

• Harmful nature of GMO

• Environmental

• Human Health

Requires that all GMOs (other than GMMs) are classified on the basis of whether or not they are HSE notifiable

GMO poses no risk to man greater than that of the parental organism

GMO poses a greater risk to man than that of the parental organism

HS

E n

otif

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The GMO (Contained use) Regulations

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Keep paperwork up-to-date

• Review when you make a change

• How?– GM Form C

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GM WASTE

• Inactivate by validated means

• Method must work under the conditions of use

• Chemical disinfection/autoclaving

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INFORMATION SOURCES

•HSE Website: http://www.hse.gov.uk/hthdir/noframes/hthdir.htm•SACGM Website: http://www.hse.gov.uk/biosafety/gmo/acgm/acgmcomp/index.htm•ACDP Website: http://www.hse.gov.uk/hthdir/noframes/agent1.pdf•GM Regs: http://www.hmso.gov.uk/si/si2000/20002831.htm•Safety Department Website: http://www3.imperial.ac.uk/safety/guidanceandadvice/biosafety

Workshop Scenario

• You enter a Containment level 2 lab to retrieve your bacterial cultures that have been growing overnight in a shaking incubator. Upon entering the lab you find that there has been a spillage of an unknown culture from one of the incubators onto the floor.

• What do you do?

Model Answer

• Switch off incubator

• Ascertain what was in the culture by referring to the labelling system or log book

• Contact relevant researcher and local safety officer

• Refer to spillage SOP

• Wear appropriate PPE

• Use disinfectants at the specified conc and contact time.

• Ensure safe disposal of contaminated material.

• Complete College DO form

• If Class 2 GM material Safety Dept must notify HSE. Further investigations required.