Recombinant DNA Training for Research Group Members Recombinant DNA Training for Research Group Members

How to Use this Training TemplateHow to Use this Training Template

Principal Investigators (PIs) whose research Principal Investigators (PIs) whose research involves recombinant DNA are required by NIH involves recombinant DNA are required by NIH to provide their research group members to provide their research group members training on lab-specific hazards and emergency training on lab-specific hazards and emergency procedures. The Berkeley Biosafety Program(procedures. The Berkeley Biosafety Program(bso@berkeley.edu) has prepared this ) has prepared this PowerPoint presentation template to assist PIs PowerPoint presentation template to assist PIs in meeting this requirement. in meeting this requirement. Text in italics Text in italics [example] [example] indicates where the template should indicates where the template should be modified to accurately reflect your lab be modified to accurately reflect your lab hazards and procedures. hazards and procedures.

Working with Recombinant Working with Recombinant DNA in the DNA in the

_[name of PI]_ _[name of PI]_ LaboratoryLaboratory

[Name of PI] [Name of PI]

[Name of Department][Name of Department]

[Date Training Is Given][Date Training Is Given]

Basic RequirementsBasic RequirementsResearch involving recombinant DNA Research involving recombinant DNA is closely regulated by the National is closely regulated by the National Institutes of Health (NIH), and requires Institutes of Health (NIH), and requires a Biological Use Authorization (BUA) a Biological Use Authorization (BUA) approved by the Berkeley campus approved by the Berkeley campus Committee on Laboratory & Committee on Laboratory & Environmental Biosafety (CLEB). CLEB Environmental Biosafety (CLEB). CLEB works with the campus Biosafety works with the campus Biosafety Program, which can provide additional Program, which can provide additional guidance (guidance (bso@berkeley.edu))

Basic Requirements (ContBasic Requirements (Cont’’d)d)All work involving recombinant DNA – All work involving recombinant DNA – regardless of funding source -- must regardless of funding source -- must adhere to the mandatory NIH adhere to the mandatory NIH ““Guidelines for Research Involving Guidelines for Research Involving recombinant DNA Moleculesrecombinant DNA Molecules”” found found at at http://oba.od.nih.gov/rdna/nih_guidelines_oba.html

Purpose of This TrainingPurpose of This TrainingSection IV-B-7-d-(2) of the NIH GuidelinesSection IV-B-7-d-(2) of the NIH Guidelines

The NIH requires that the PI provide The NIH requires that the PI provide lab-specific biosafety training on the lab-specific biosafety training on the hazards of the specific experimental hazards of the specific experimental procedures used in their laboratory procedures used in their laboratory to members of their research group. to members of their research group. One way to document this training is One way to document this training is to record it in the minutes of a group to record it in the minutes of a group meeting, listing all participants.meeting, listing all participants.

Contents of This TrainingContents of This Training

• CLEB Administrative RequirementsCLEB Administrative Requirements• What are the NIH Guidelines?What are the NIH Guidelines?• Role of the Investigator/Lab StaffRole of the Investigator/Lab Staff• Description of the Experiment-BUADescription of the Experiment-BUA

– The Agents UsedThe Agents Used– The TemplateThe Template

• Risk Assessment/Biosafety LevelRisk Assessment/Biosafety Level– Good Work PracticesGood Work Practices– Personal Protective EquipmentPersonal Protective Equipment– Engineering ControlsEngineering Controls

• Waste Disposal/DecontaminationWaste Disposal/Decontamination• Emergency ProceduresEmergency Procedures

CLEB Administrative CLEB Administrative RequirementsRequirements

• CLEB requires the following from CLEB requires the following from every PI approved to use recombinant every PI approved to use recombinant DNA:DNA:– Train all research group members on lab-Train all research group members on lab-

specific hazards and proceduresspecific hazards and procedures– Report incidents involving recombinant Report incidents involving recombinant

DNA immediately to EH&SDNA immediately to EH&S– Submit a BUA amendment for any change Submit a BUA amendment for any change

in personnel or Scope of Workin personnel or Scope of Work

What are the NIH What are the NIH Guidelines?Guidelines?Key Sections of the NIH GuidelinesKey Sections of the NIH Guidelines

• Appendix B-List of AgentsAppendix B-List of Agents• Section III-Types of Experiments involving Section III-Types of Experiments involving

recombinant DNArecombinant DNA• Appendix G-Level of ContainmentAppendix G-Level of Containment• Appendix K-Large ScaleAppendix K-Large Scale• Appendix M-Gene Transfer/Human Appendix M-Gene Transfer/Human

Subjects Subjects • Appendix P-Transgenic PlantsAppendix P-Transgenic Plants• Appendix Q-Transgenic AnimalsAppendix Q-Transgenic Animals• Section IV-Responsibilities of PI, IBC Section IV-Responsibilities of PI, IBC

PI RESPONSIBILITIESPI RESPONSIBILITIESSection IV-B-7: ROLE OF THE PISection IV-B-7: ROLE OF THE PI

On behalf of the institution, the Principal On behalf of the institution, the Principal Investigator is responsible for full compliance Investigator is responsible for full compliance with the with the NIH Guidelines NIH Guidelines for any recombinant for any recombinant DNA research. DNA research.

The PI must:The PI must:• Not initiate or modify recombinant DNA research Not initiate or modify recombinant DNA research

which requires CLEB approval until that research has which requires CLEB approval until that research has been approved by CLEB and has met all other been approved by CLEB and has met all other requirements of the requirements of the NIH Guidelines.NIH Guidelines.

• Report any significant problems, violations of the Report any significant problems, violations of the NIH NIH GuidelinesGuidelines, or any significant research-related , or any significant research-related accidents and illnesses to the Biological Safety Officer accidents and illnesses to the Biological Safety Officer and CLEB.and CLEB.

OTHER PI RESPONSIBILITIESOTHER PI RESPONSIBILITIES

ResponsibilityResponsibility NIH NIH Guideline ReferenceGuideline Reference

• Train Lab Staff (Micro technique)Train Lab Staff (Micro technique) Section IV-B-7-a, dSection IV-B-7-a, d• Ship Agents SafelyShip Agents Safely Section IV-B-7-aSection IV-B-7-a• Follow Emergency PlansFollow Emergency Plans Section IV-B-7-aSection IV-B-7-a• Notify NIH of novel host/vectorsNotify NIH of novel host/vectors Section IV-B-7-bSection IV-B-7-b• Update ProtocolUpdate Protocol Section IV-B-7-cSection IV-B-7-c• Notify IBC/CLEB of incident Notify IBC/CLEB of incident Section IV-B-7-eSection IV-B-7-e• Make Protocol AvailableMake Protocol Available Section IV-B-7-dSection IV-B-7-d• Offer Medical SurveillanceOffer Medical Surveillance Section IV-B-7-dSection IV-B-7-d• Supervise Lab MembersSupervise Lab Members Section IV-B-7-eSection IV-B-7-e• Maintain Safety EquipmentMaintain Safety Equipment Section IV-B-7-eSection IV-B-7-e

Description of the Experiment-Description of the Experiment-BUABUAThe Scope of WorkThe Scope of Work• [Cut and paste scope of work narrative from your BUA] [Cut and paste scope of work narrative from your BUA]

Description of the Experiment-Description of the Experiment-BUABUAThe Agents UsedThe Agents Used

[List the agents, recombinant DNA host/vector [List the agents, recombinant DNA host/vector systems, human cell lines, packaging cell lines, and systems, human cell lines, packaging cell lines, and clinical specimens used in the experiment. Include clinical specimens used in the experiment. Include all etiological agents.]all etiological agents.]

Description of the Experiment-Description of the Experiment-BUABUAThe Risks That May Affect Containment The Risks That May Affect Containment Level Level

• These are some of the common These are some of the common experimental risks associated with our experimental risks associated with our research: research: [modify list as appropriate][modify list as appropriate]– Route(s) of exposureRoute(s) of exposure– volumes of culturevolumes of culture– aerosol generating proceduresaerosol generating procedures– Procedures involving sharpsProcedures involving sharps– high concentrations high concentrations – the use of vectors and vulnerable animalsthe use of vectors and vulnerable animals– inserted genetic sequences that increase virulenceinserted genetic sequences that increase virulence– the use of drug/antibiotic resistant strainsthe use of drug/antibiotic resistant strains

Description of the Experiment-Description of the Experiment-BUABUAThe Risks That May Affect Containment The Risks That May Affect Containment Level Level

•These are the symptoms of exposure These are the symptoms of exposure with the agents we use:with the agents we use:

– [List symptoms of exposure][List symptoms of exposure]

Our Recombinant DNA Experiment Our Recombinant DNA Experiment TemplateTemplate[Cut and paste from BUA form:][Cut and paste from BUA form:]

• Agent Characteristics: Agent Characteristics:

• Routes of Exposure: Routes of Exposure:

• HostHost: :

• VectorVector: :

• Nature of inserted sequencesNature of inserted sequences: :

• Source of inserted sequencesSource of inserted sequences: :

• Types of manipulation: Types of manipulation:

• Attempt to express foreign gene:Attempt to express foreign gene:

• Protein produced: Protein produced:

• Containment: Containment:

• Section of Guidelines: Section of Guidelines:

Risk Assessment/Biosafety Risk Assessment/Biosafety LevelLevelGood Work Practices Good Work Practices [Modify list as [Modify list as appropriate]appropriate]

• Wash Hands FrequentlyWash Hands Frequently

• Use Personal Protective Equipment Use Personal Protective Equipment and Engineering Controlsand Engineering Controls

• Minimize Use of SharpsMinimize Use of Sharps

• Do Not Eat, Drink, or Smoke in the Do Not Eat, Drink, or Smoke in the lablab

• Disinfect Work AreasDisinfect Work Areas

Risk Assessment/Biosafety Risk Assessment/Biosafety Level Level Personal Protective Equipment Personal Protective Equipment [Modify list as appropriate][Modify list as appropriate]

• The following personal protective The following personal protective equipment must be worn when equipment must be worn when handling agents in the tissue culture handling agents in the tissue culture room and the biosafety cabinet:room and the biosafety cabinet:– Latex or nitrile glovesLatex or nitrile gloves– Lab coat Lab coat – Safety glassesSafety glasses

Risk Assessment/Biosafety Risk Assessment/Biosafety Level Level Engineering Controls Engineering Controls [Modify list as [Modify list as appropriate]appropriate]

• The following engineering controls must be The following engineering controls must be used whenever handling agents in the used whenever handling agents in the tissue culture room:tissue culture room:– Biosafety cabinetBiosafety cabinet– Sink for Hand WashingSink for Hand Washing– Mechanical pipettesMechanical pipettes– Sealed vialsSealed vials– Sharps containers/Broken Glass boxSharps containers/Broken Glass box– Eyewash/deluge shower (use only in an Eyewash/deluge shower (use only in an

emergency)emergency)

Waste Waste Disposal/DecontaminationDisposal/Decontamination[Modify list as appropriate][Modify list as appropriate]• Liquid waste will be treated with 10% Liquid waste will be treated with 10%

household bleach and allowed to set for household bleach and allowed to set for 20 minutes and drain-disposed. Solid red 20 minutes and drain-disposed. Solid red bag wastes will be placed in a red-bag bag wastes will be placed in a red-bag lined bin and transferred to the _________ lined bin and transferred to the _________ Building, Room______ medical waste Building, Room______ medical waste storage area for pick up and disposal.storage area for pick up and disposal.

• Work areas must be disinfected with Work areas must be disinfected with dilute bleach and 70% ethanol.dilute bleach and 70% ethanol.

Emergency Procedures Emergency Procedures [modify as [modify as

appropriate]appropriate]

• In the event of a laboratory accident:In the event of a laboratory accident:– Tend to injured personTend to injured person– Notify the PI or the lab supervisor; Notify the PI or the lab supervisor;

inform the PI IMMEDIATELY if inform the PI IMMEDIATELY if recombinant DNA was involved so recombinant DNA was involved so that CLEB and the Biosafety Officer that CLEB and the Biosafety Officer can be notified within 24 hrs. can be notified within 24 hrs.

– Initiate clean up procedures.Initiate clean up procedures.– Notify EHS, if the PI is not available, Notify EHS, if the PI is not available,

642-3073 or 642-3333 (After Hours)642-3073 or 642-3333 (After Hours)– Seek medical attention at Tang or Seek medical attention at Tang or

the Alta Bates ER at 2450 Ashby the Alta Bates ER at 2450 Ashby Avenue after hours. Avenue after hours.

ConclusionConclusion

This training is designed to inform you of This training is designed to inform you of the NIH requirements regarding use of the NIH requirements regarding use of recombinant DNA, and our labrecombinant DNA, and our lab’’s s implementation of the NIH requirements. implementation of the NIH requirements. All recombinant DNA experiments require All recombinant DNA experiments require BUA approval by CLEB and any changes BUA approval by CLEB and any changes require an approved amendment to the require an approved amendment to the BUA.BUA. If you have any questions on this If you have any questions on this training, let me know, and I will contact training, let me know, and I will contact the Biosafety Officer for assistance. the Biosafety Officer for assistance.