1. Foundation Training in Biological Safety

2. Module 4 Genetically modified organisms

3. Why GM?

5. Page 5 Gene of interest (insert) Vector Host Protein of interest What is GM? (e.g. bacterial expression system)

8. Page 8 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

9. Not GM

11. 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

12. Page 12 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

14. Page 14 Gene of interest (insert) Vector Host Protein of interest Example of GM (bacterial expression system)

15. Page 15 Risks HOST –How pathogenic? –Virulence and infectivity? –Are toxins produced? –Allergenic factors? –Seriousness of exposure? –Survivability outside lab?

16. E. coli K12, the prokaryotic wimp

17. Page 17 RISKS VECTORS –Consider: – Pathogenicity –Transferability of the vector: –Only want vector to transfer target host, achieved by modifying how a vector transfers

19. Page 19 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?

20. THE GM (CONTAINED USE) REGULATIONS Requires that all GMMs are classified in one of four Classes Class 1Class 2Class 3Class 4 Increasing hazard to human health or the environment 1234 Containment Level Classification of GMM determined by Containment Level required

21. Page 21

22. Page 22 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

25. Page 25 RISK ASSESSMENT APPROACH FOR GM ANIMALS AND PLANTS Harmful nature of GMO Environmental Human Health

26. 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 HSE notifiable The GMO (Contained use) Regulations

27. Page 27 Keep paperwork up-to-date Review when you make a change How? –GM Form C

28. Page 28 GM WASTE Inactivate by validated means Method must work under the conditions of use Chemical disinfection/autoclaving

29. Page 29 INFORMATION SOURCES HSE Website: http://www.hse.gov.uk/hthdir/noframes/hthdir.htm SACGM Website: http://www.hse.gov.uk/biosafety/gmo/acgm/acgmcom p/index.htm ACDP Website: http://www.hse.gov.uk/hthdir/noframes/agent1.pdf GM Regs: http://www.hmso.gov.uk/si/si2000/20002831.htm http://www.hmso.gov.uk/si/si2000/20002831.htm Safety Department Website: http://www3.imperial.ac.uk/safety/guidanceandadvice /biosafety http://www3.imperial.ac.uk/safety/guidanceandadvice /biosafety

32. 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?

33. 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.