Download BUA Application Details (BU041135-03)

BUA Application Details (BU041135-03)
Summary
Principal Investigator :
PI Department :
Laboratory Supervisor :
Title :
Status :
Expiration date :
OEH&S Specialist :
Application Type :
NIH Level :
McManus, Michael T.
Diabetes Center
Michael T. McManus
Lentiviral RNAi Core
Approved
04/01/2018
Herminigildo Briones
Continuation
Section III-D-1
Study involves
Recombinant DNA materials or technology: Yes
Infectious Agents (IA): No
Bloodborne Pathogens (BBP): Yes
Biological Toxins: No
Select Agents : No
Generate Genetically-Modified Animals: No
Animals: No
Shipping of biological materials: Yes
Describe the goals of your research and an overview of the experimental design to attain these goals.
Among the 3 billion base pairs of human genome sequence, there are at least 30-40 thousand protein-encoding genes, but the
function of at least half of them remains unknown. The use of lentiviral vectors and other viral agents (including MMLV,
MSCV, and AAV) are powerful tools for systematically deciphering the functions and interactions of these thousands of genes.
Their ability to transduce both dividing and non-dividing cell types allows for their application in both overexpression and
RNAi experiments.
The overall goal of our research is to provide high titer virus to researchers in the Immunology and Microbiology Departments
and the Diabetes Center and members of the UCSF community. This involves:
•
Packaging of both shRNA, gRNA, and cDNA expressing self-inactivating lentiviral vectors, MMLV, AAV, and other
viral agents upon request.
•
Titration of produced virus by flow cytometry.
•
Providing a resource for protocols and safety regarding work with infectious reagents.
All labs for which we will provide lentiviral or other vectors will have active BUAs that include these biomaterials.
No significant changes have occurred since our last BUA.
Project Number
BU041135-03
BU041135-02C
BU041135-02B
BU041135-02A
BU041135-02
BU041135-01
Approval Type
Continuation
Modification
Modification
Modification
Continuation
New Approval
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Application History
Approval Date
03/10/2015
01/17/2014
06/14/2013
08/15/2012
02/15/2012
Expiration Date
04/01/2018
03/01/2015
03/01/2015
03/01/2015
03/01/2015
12/01/2011
1
Funding
Type:
Funding Agency/Sponsor
Grant or Contract Number
Applicant comments:
We are funded by recharge.
Associated Authorizations/Protocols
Regulated Materials
Approval Number
Legacy Number
Status
Expiration Date
Recombinant DNA Research
I am transferring genetic material into a microbiological vector or any organism in order to clone the
gene, create a stable genetic alteration, obtain the expression product(s) of the gene(s), or silence/knockdown the target gene(s). The genetic material is not from the same species of organism or vector that is
receiving it.
I am introducing foreign genetic material using a vectorless system (e.g. electroporation, lipofection).
The genetic material being cloned or expressed is from a Risk Group (RG) 3 agent.
The genetic material being cloned or expressed is from a Risk Group (RG) 4 agent.
The genetic material being cloned or expressed consists of more than two-thirds of the genome of a RG 1
or RG 2 Eukaryotic virus.
The genetic material being cloned or expressed encodes a known or reasonably suspected Oncogene or
Teratogen.
The vector I am using for introduction of foreign genetic material into the host is, or is derived from, a RG
3 agent.
The vector I am using for introduction of foreign genetic material into the host is a RG 1 or RG 2
Eukaryotic virus which contains more than two-thirds of its viral genome, or more than two-thirds of
another Eukaryotic virus' genome.
The vector I am using for introduction of foreign genetic material into the host is being used in
conjunction with a helper virus.
The host into which I am introducing foreign genetic material is a cell or organism other than E. coli
strain K12; S. cerevisiae, S. uvarum, B. Subtilis, or B. licheniformis?
I will be making more than 10 liters, in one container, of any culture.
I am using synthetic nucleic acid molecules, including those that are chemically or otherwise modified but
can base pair with naturally occurring nucleic acid molecules and/or cells, organisms, and viruses
containing such molecules.
The gene(s) I am transferring into a microorganism encode for antibiotic resistance that the organism
would not acquire naturally, and the resistance could compromise the use of the antibiotic to control
disease caused by the receiving organism in human medicine, veterinary medicine or agriculture.
The gene(s) I am transferring into a microorganism for biosynthesis of a toxin lethal to vertebrates with
an LD50 of less than 100 nanograms per kilogram body weight (100ng/Kg).
I am transferring recombinant DNA, or DNA or RNA derived from recombinant DNA into one or more
human research subjects.
No
Yes
No
No
No
No
No
No
No
Yes
No
No
No
No
No
Describe experimental procedures that will involve the rDNA listed here. You should tell the IBC (Institutional
Biosafety Committee) not only what you will do but also how you will do it. Step-by-step descriptions of procedures are
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not necessary, but focus should be placed on procedural steps where personnel are vulnerable to exposure to potentially
hazardous recombinant materials.
We will use lipid transient transfection methods to introduce multiple DNA plasmids containing the different viral genes
necessary for propagating virus along with the shRNA/gRNA/gene of interest into HEK 293t cells. The viral genomes are
broken into separate plasmids and several replication genes have been removed to ensure viral particles are replication
incompetent. We will harvest and concentrate virus and other viral agents 48-72 hours post-transfection using syringes, filters,
and ultracentrifugation methods. To titer the virus we will infect the appropriate host cell (e.g. HEK 293t cells for lentivirus)
with a dilution of virus and then use flow cytometry to calculate the titer values. We will not be doing actual gene
insertions/creation of gene constructs for other labs, or any other rDNA procedures (i.e. siRNA or other gene targeting, sitedirected mutagenesis, etc.).
Please note that the viral vectors listed in the Vectors table are produced by the Core facility for researchers. The genes listed in
the "Genes" table are those potentially packaged, and the hosts listed in the "Hosts" table include the hosts that the vectors can
infect.
Vectors
Vector Name
Vector Type
Risk Group
Amphotropic
Used in Animals
1
Replication
Incompetent
Yes
Adeno-associated
Virus (AAV)
Avian Retrovirus
Lentivirus
Moloney Murine
Leukemia Virus
(MMLV)
various
Viral
Yes
Yes
Viral
Viral
Viral
1
2
2
Yes
Yes
Yes
No
Yes
Yes
No
No
No
Plasmid
1
No
No
No
Vector Name: Adeno-associated Virus (AAV)
Risk Group: 1
Vector Type: Viral
Replication incompetent: Yes
Causes human disease: No
Ecotropic: No
Amphotropic: Yes
Used in Animals: Yes
Please indicate which generation of this viral vector you will use.
We will provide serotypes AAV1-6, and primarily package serotype 2 (the most commonly used AAV).
If you derive a new vector from wild-type virus, or similarly use an early generation vector (for example, first
generation), please describe the method by which you will verify replication incompetence. Specify the frequency of
testing.
N/A
Provide a detailed description of this vector and safety features of its design (such as which generation of viral vector,
gene deletions, etc.):
AAV are infectious human viruses with no known disease association. Replication of AAV has an extremely low efficiency
without the presence of helper virus (such as adenovirus). Similar to retrovirus such as HIV, packaging recombinant virus
entails separating the replication and capsid genes and provided them transiently in trans (in a pRep/Cap plasmid), where only
the 2 inverted terminal repeats of AAV genome are left and packaged into virion; hence, the likelihood for a recombinant AAV
to convert to a replicative competency is highly improbably and some consider theoretically impossible. Recombinant AAV
constructs in which the transgene does not encode either a potentially tumorigenic gene product or a toxin molecule and are
produced in the absence of a helper virus is in most cases be handled at BSL1.
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Vector Name: Avian Retrovirus
Risk Group: 1
Vector Type: Viral
Replication incompetent: Yes
Causes human disease: No
Ecotropic: Yes
Amphotropic: No
Used in Animals: No
Please indicate which generation of this viral vector you will use.
If you derive a new vector from wild-type virus, or similarly use an early generation vector (for example, first
generation), please describe the method by which you will verify replication incompetence. Specify the frequency of
testing.
Provide a detailed description of this vector and safety features of its design (such as which generation of viral vector,
gene deletions, etc.):
Retroviral infection requires the specific interaction between the envelope glycoprotein on the surface of the virus and the
cognate receptor on the surface of the cell. The ASLV family of viruses has five primary envelope types: A, B, C, D, E. These
recognize three distinct cellular receptors: A, C, and B/D/E. In order for the virus to be propagated, your cells must have the
appropriate receptor and cannot be sequentially infected with two ASLVs expressing the same envelope. Mammalian cells do
not have functional receptors for the standard ASLV envelopes on their surface.
Vector Name: Lentivirus
Risk Group: 2
Vector Type: Viral
Replication incompetent: Yes
Causes human disease: Yes
Ecotropic: No
Amphotropic: Yes
Used in Animals: No
Please indicate which generation of this viral vector you will use.
3rd generation
If you derive a new vector from wild-type virus, or similarly use an early generation vector (for example, first
generation), please describe the method by which you will verify replication incompetence. Specify the frequency of
testing.
N/A
Provide an explanation of the hazards to humans and describe the possible modes of transmission:
Lentivirus has potential to cause human disease if there were an exposure. We have placed a page describing safety
considerations at the viracore website (http://viracore.ucsf.edu/content/biosafety).
Provide a detailed description of this vector and safety features of its design (such as which generation of viral vector,
gene deletions, etc.):
Only "third generation" lentiviral vectors, such as the pSico or LentiLox vectors available from the UCSF Sandler Lentiviral
RNAi Core. These systems separate vector and packaging functions onto three or four or more plasmids and they include
additional safety features (e.g., they do not encode Tat, which is essential for replication of wild-type HIV-1).
Vector Name:
Risk Group:
Vector Type:
Replication incompetent:
Causes human disease:
Ecotropic:
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Moloney Murine Leukemia Virus (MMLV)
2
Viral
Yes
Yes
Yes
4
Amphotropic: Yes
Used in Animals: No
Please indicate which generation of this viral vector you will use.
MMLV (and other murine retroviruses) can be packaged as ecotropic or amphotropic. Here we primarily package the
ecotropic as it offers a specificity for murine cells. In these cases, packaging can be performed safely with pCL-ECO (a first
generation murine packaging vector). In very rare cases we may package MMLV with amphotropic coats (I think we needed
to do this once or twice in the last 8 years). In these rare cases we will use the replication safety assay to monitor for potential
conversion.
If you derive a new vector from wild-type virus, or similarly use an early generation vector (for example, first
generation), please describe the method by which you will verify replication incompetence. Specify the frequency of
testing.
N/A
Provide an explanation of the hazards to humans and describe the possible modes of transmission:
Non-replicating MMLV retroviruses, if packaged with amphotropic envelope or VSV-G envelope,can infect human cells and
integrate into the cell's genome. These viruses are non-replicating, so cannot spread after the initial infection. Nonetheless, we
will use a replication safety assay if such a virus is packaged with the amphotropic coats.
These are described in the UCSF Sandler Lentiviral RNAi Core’s MMLV MSDS SOP, found on our Core website:
http://viracore.ucsf.edu/biosafety, and appended. The vectors used will generate only non-replicating viral particles.
Provide a detailed description of this vector and safety features of its design (such as which generation of viral vector,
gene deletions, etc.):
We may employ the non-replicating pMX MMLV retroviral backbone vector which has been used by many labs for more than
a decade. The non-replicating pMX-based retroviral vector and related pMIGR1 retroviral vector are described in Exp
Hematol. 31:1007, 2003 and Blood 92:3780, 1998. The enhanced green fluorescent protein (eGFP)-expressing retroviral
vector, MigR1, was constructed by three-way ligation with the 5-kb EcoRI-Sal I cleaved MSCV2.2, the 0.5-kb IRES
containing EcoRI-Nco I fragment from pCITE, and the 0.8-kb Nco I-Sal I fragment containing a GFP with solubility and red
shift mutations. The pMX vector harbors 5′ long terminal repeat (LTR) and the extended packaging signal derived from MFG
followed by a multi-cloning site (MSC) suitable for cDNA library construction and 3′ LTR of MMLV. The pMX and pMIG
vectors are commonly packaged in Phoenix-Eco or Phoenix-Amp cell line, which are 293T cells stably transfected with the
MMLV gag and pol genes on one plasmid vector and the envelope (ecotropic or amphotrophic) gene on a separate plasmid
vector. Having these elements on separate plasmids, with the gag and pol stably integrated into the 293T packaging cells
guards against the creation of replicating viruses.
Vector Name: various
Risk Group: 1
Vector Type: Plasmid
Replication incompetent: No
Causes human disease: No
Ecotropic: No
Amphotropic: No
Used in Animals: No
Provide a detailed description of this vector and safety features of its design (such as which generation of viral vector,
gene deletions, etc.):
Genes
Gene (Family)
Name
All genes
DNA Sources
Immunogenic
(Proto)Oncogenic
Toxigenic
Pathogenic
Human and mouse
No
Yes
No
No
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GFP derivatives
Sea Pansy
No
No
No
No
All genes
Human and mouse
2
Yes
We make whole-genome cDNA and RNA libraries in which
every single mouse and human gene are expressed. This
constitutes broad families of genes that include adapter proteins,
kinases, or phosphatases, signalling molecules, transcription
factors, etc or mutants thereof. Ideally all known gene ontologies
are represented in our libraries since we wish to be
comprehensive in our studies. Similarly we will generate whole
genome libraries that contain gene interfering agents targeting
every known mouse and human gene.
Adverse Immune Reaction: No
(Proto) Oncogenic: Yes
Toxigenic: No
Enhances Pathogenicity: No
Pathogenic to Humans: No
Teratogenic: No
Not sure: Yes
None: No
Associated vectors: Adeno-associated Virus (AAV)/Viral, Avian Retrovirus/Viral,
Lentivirus/Viral, Moloney Murine Leukemia Virus
(MMLV)/Viral, various/Plasmid
Describe the nature of cellular activity that will result if this gene is expressed or knocked down.
All known classes of genes annotated in the human and mouse genome. Therefore genes of potential health concerns are
targeted by this library. The most obvious gene group that may be of potential concern are the tumor supressor families, in
which RNAi knockdown- if accomplished in a cell containing a 'second hit' or driver mutation, may yield cells that proliferate
in an oncogenic manner. We address safety considerations below.
Provide an assessment of the hazardous potential of cloning, expressing or inhibiting the target of these DNA/RNA
segments that encode substances that are immunogenic, (proto)oncogenic, teratogenic, toxigenic or enhance
pathogenicity:
Over-expression of some genes or silencing of tumor suppressor genes can be proto-oncogenic.
Gene or Gene Family Name:
Gene Biological Sources:
Highest Risk Group of Sources:
Expressed as RNA/Protein:
Indicate which RNA or protein will be produced:
Virus work is performed in a biosafety cabinet and contact with the researcher is not expected.
Gene or Gene Family Name:
Gene Biological Sources:
Highest Risk Group of Sources:
Expressed as RNA/Protein:
Indicate which RNA or protein will be produced:
Adverse Immune Reaction:
(Proto) Oncogenic:
Toxigenic:
Enhances Pathogenicity:
Pathogenic to Humans:
Teratogenic:
Not sure:
BU041135-Y03-V21-2015-03-19-Final.doc
GFP derivatives
Sea Pansy
1
Yes
We will express RNA which will be translated as a fluorescent
protein
No
No
No
No
No
No
No
6
None: Yes
Associated vectors: Adeno-associated Virus (AAV)/Viral, Avian Retrovirus/Viral,
Lentivirus/Viral, Moloney Murine Leukemia Virus
(MMLV)/Viral, various/Plasmid
Describe the nature of cellular activity that will result if this gene is expressed or knocked down.
This is an ectopically expressed gene and overexpression does not appear to be broadly toxic in a wide variety of cell lines and
organisms.
Provide an assessment of the hazardous potential of cloning, expressing or inhibiting the target of these DNA/RNA
segments that encode substances that are immunogenic, (proto)oncogenic, teratogenic, toxigenic or enhance
pathogenicity:
No known hazards when working with these types of genes.
Hosts
Host Name
HEK Cells
human established cell lines
Infectious Agent
No
No
Host Name:
Risk Group:
Infectious Agent:
Bloodborne Pathogen:
Host details (if applicable):
Bloodborne Pathogen
Yes
Yes
HEK Cells
2
No
Yes
Vector-Gene Construct(s) for HEK Cells
Vectors \ Genes:
Adeno-associated Virus (AAV)
Avian Retrovirus
Lentivirus
Moloney Murine Leukemia Virus
(MMLV)
various
All genes
Yes
Yes
Yes
Yes
GFP derivatives
Yes
Yes
Yes
Yes
Yes
Yes
Host Name:
Risk Group:
Infectious Agent:
Bloodborne Pathogen:
Host details (if applicable):
human established cell lines
2
No
Yes
In the case of Avian Retrovirus infection, this cell line has been
modified to express the receptor that allows for adherence.
Vector-Gene Construct(s) for human established cell lines
Vectors \ Genes:
Adeno-associated Virus (AAV)
Avian Retrovirus
Lentivirus
Moloney Murine Leukemia Virus
(MMLV)
various
All genes
Yes
Yes
Yes
Yes
GFP derivatives
Yes
Yes
Yes
Yes
Yes
Yes
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rDNA Safety
Confirm that, in case of accidental exposure to recombinant materials, you will follow this SOP:
Personnel who are exposed to recombinant materials will immediately wash the site of exposure (wound, eye, mucosa, etc.)
with copious amounts of water for 15 minutes and then immediately call the Needlestick and Exposure Hotline at 415-3537842 for all UCSF campuses and affiliated institutions. Important: Report exposure to Biosafety Officer (415-514-2824) within
72 hours.
If the SOP cannot be followed, describe how exposure will be managed:
SOP will be followed
Describe methods by which experiment(s) will be conducted safely:
We will use Universal Precautions and work at Biosafety Level 2 containment. We will avoid, when possible, use of needles
or sharps since this is the greatest risk. 1) the Personal Protective Equipment (PPE) that lab personnel will wear (i.e. gloves,
lab coats, eye protection (if needed), etc.), 2) Waste decontamination (final dilution of 10% bleach is used to decontaminate
liquid waste for 30 minutes before drain disposal, solid rDNA waste sent out as biohazardous waste), and 3) use of a certified
Class II Biosafety cabinet for rDNA procedures. There will be no work with Diphtheria Toxin Receptor or Diphtheria Toxin.
If these constructs were accidentally released outside of the laboratory, will there be significant implications to the
environment? If 'Yes', please describe:
No.
Bloodborne Pathogens
Human tissue, including bones or teeth:
A primary cell line or cells derived directly from human tissue:
Human blood, blood products, body fluids, other potentially infectious materials (OPIM):
Human established cell lines:
BBP Material
Human established cell lines
No
No
No
Yes
Used in Animals
No
Potential Bloodborne Pathogen Exposure: Human established cell lines
Risk Group: 2
Used in Animals: No
Describe the specific human materials you are using here (e.g. liver, MCF, prostate cancer cells, etc.) and how you will
use them:
Numerous established human cell lines. These will be used to generate lentivirus and for flow cytometry. They will be
handled as BLS2 using appropriate BBP procedure.
Describe the experimental procedures where personnel are vulnerable to exposure to potential bloodborne pathogens.
Describe your source(s) for human materials, how they are packaged and transported to your lab, and provide the
CHR approval number for their use or a statement that the CHR has exempted this work. Add CHR approval number
to the 'Associated Auth/Protocol' page.
Established cell line derived from human tissue, such as 293T. These cells were immortalized decades ago and used in a large
number of labs and are available from many sources. We have obtained these from the ATCC and UCSF Cell Culture Facilityvia cryovials. Cells will be transported with a leak-proof secondary container.
Describe the hazardous potential and the risk to humans from accidental exposure to BBP (e.g. needle stick/sharps
injury; exposure to eyes, mucosa, wounds by aerosol, splash or spill, etc):
It should be noted that handling human cells entails risk of exposure to Hepatitis B, Hepatitis C, and/or HIV. Technically the
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pathological transfer of cell lines could occur in a needle stick/sharps injury, exposure to eyes, mucosa, wounds by aerosol,
splash or spill to an immunocompromised individual. In addition, if a contaminating agent was present, the individual need not
be immunocompromised.
Confirm that you will adhere to the Bloodborne Pathogens Standard, use Universal Precautions and work at Biosafety
Level 2 containment, including:
- Use certified biosafety cabinet for all manipulations of BBP
- Require use of appropriate PPE, i.e., lab coats or gowns, gloves, eye protection, etc.
- Ensure that I and all personnel listed on the BUA will complete annual online BBP training and triennial online Biosafety
training
- Offer Hepatitis B vaccination to all personnel listed on the BUA
- Ensure that all vaccination records and declination forms will be maintained by the Occupational Health Service
If any of the requirements cannot be met, describe alternative or additional methods by which experiments will be
conducted safely:
The above requirements will be met.
Confirm that, in case of accidental exposure to biohazardous materials, you will follow this SOP:
Personnel who are exposed to biohazardous materials will immediately wash the site of exposure (wound, eye, mucosa, etc.)
with copious amounts of water for 15 minutes and then immediately call the Needlestick and Exposure Hotline at 415-3537842 for all UCSF campuses, except SFGH and VA Medical Center call 469-4411.
If the SOP will not be followed, describe how exposure will be managed:
SOP will be followed
Standard Procedures
Decontamination Procedures: I will use 0.5% sodium hypochlorite (a 1:10 dilution of household bleach) to
decontaminate equipment and work surfaces. In locations where bleach would cause corrosion, I will
decontaminate with 70% ethanol if effective against the agent(s) used.
Local Shipping and Transport of Infectious Materials: I will follow the procedures outlined in the UCSF
Infectious Agents Shipping and Transport Policy. This flyer must be posted on orange paper in BSL 2
location(s) in the laboratory. Pre-printed flyer(s) are available from OEH&S Specialists. In brief, all
infectious materials transported to and from my laboratory will be enclosed in a primary container with
sealed lid or top, which will then be enclosed in a secondary leak-proof, non-breakable container (such as
a Coleman cooler) appropriately labeled with the biohazard symbol. Any specimens transported to and
from off-campus satellite facilities will be escorted by a responsible lab employee.
Storage: All infectious materials to be stored will be clearly labeled with the universal biohazard symbol,
as will the storage space (e.g. freezer, refrigerator, liquid nitrogen tank).
Bloodborne Pathogens: I will follow the Standard Precautions and procedures outlined in the Bloodborne
Pathogens training.
Human Organ and Cell Culture: If I am using human organ or cell cultures (primary cultures, cell
strains, cell lines), I will follow the procedures outlined in the UCSF Cell Culture Guidelines. In addition, I
will handle all such cultures under BSL2 conditions and in accordance with the Bloodborne Pathogen
Standard unless the Institutional Biosafety Committee (IBC) has specifically approved a lower standard of
containment.
Yes
Yes
Yes
Yes
Yes
If you will use alternative procedures for any of the above, provide a brief description of the procedures that you will
follow and justification for deviating from the standard procedure.
Standard Procedures II
Biohazardous Spills: I will follow the UCSF Biological Spill Emergency Procedures. This flyer must be Yes
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posted on orange paper in BSL 2 location(s) in the laboratory. Pre-printed flyer(s) are available from
OEH&S Specialists. In brief, in case of a spill or accident involving employee exposure, I will contact the
24-hour Needlestick and Exposure Hotline at 353-7842 at all campuses except for Bloodborne Pathogens
exposures at SFGH and VA Medical Center, call 469-4411.
Disposal: I will follow the UCSF Medical Waste Policy and Procedures flyer: This flyer must be posted on Yes
orange paper in BSL 2 location(s) in the laboratory. Pre-printed flyer(s) are available from OEH&S
Specialists.
In brief:
- Liquid biohazardous waste must be decontaminated by mixing one volume of undiluted bleach with nine volumes of liquid
biohazardous waste (final dilution of 1:10) for 30 minutes. This can be drain-disposed if not mixed with chemical or
radioactive material.
- Solid biohazardous waste must be disposed in red biohazardous waste bags, which must be placed inside a hard-sided waste
container with a tight-fitting lid at all times when container is not in active use.
If you will use alternative procedures for any of the above, provide a brief description of the procedures that you will
follow and justification for deviating from the standard procedure.
Safe Sharps
For the procedures in your lab that use needles or blades, if the sharp cannot be replaced with a blunt (for example, a
needle replaced with a cannula), determine whether the needle or blade can be replaced with a safer version.
I am unable to adopt a safer sharp for legitimate reasons as provided in the BBP Standard.
Specify the sharps and/or safe sharps you will use and provide a brief description of all procedures involving sharps.
Aside from glass Pasteur pipettes used in tissue culture, we do not routinely use sharps. We will opt to use safe shares
whenever possible; for example, we use scissors.
We sometimes have the need to excise slices of gels for purifying DNA fragments. This procedure require a long-edged flat
surface with a very thin razor type material. Although double-sided razors offer the ideal material, we have opted for singleedge razors for safety reasons. In terms of safety performance, and objective product evaluation criteria, the we do not believe
the available alternative safe devices are more effective in preventing exposure incidents.
Confirm that you will follow the UCSF Proper Handling and Disposal Procedures for Sharps flyer. This flyer must be
posted on orange paper in your laboratory. Pre-printed flyer(s) are available from OEH&S
If you will use alternative handling procedures, provide a brief description of the procedures that you will follow and
justification for deviating from the standard procedure.
We will follow the UCSF Proper Handling and Disposal Procedures for Sharps.
Aerosol Safety
Do any experimental procedures or use of equipment (such as cell sorters, centrifuges, shakers, sonicators,
etc.) associated with your research produce aerosols?
Cell Sorters:
Centrifuges
Sonicators
Robotic Pipetters:
Vortexers:
Containment of Aerosols and/or Splashes. Confirm that you will follow the Standard Procedures for
Containment of Aerosols and/or Splashes: All manipulations having a potential for generating aerosols
(e.g., homogenization, centrifugation, sonification) will be conducted in a properly certified biosafety
BU041135-Y03-V21-2015-03-19-Final.doc
Yes
Yes
Yes
No
Yes
No
Yes
10
cabinet or in a centrifuge equipped with sealed rotor heads or safety cups. Screw-cap centrifuge tubes will
be no more than three-fourths filled.
If you will use additional procedures and/or equipment that may generate aerosols and/or alternative methods to
achieve containment, please describe here:
We will follow BBP Training procedures. Our FACS Array machine is located in the ViraCore and is equipped to minimize
exposure to aerosols as the samples are analyzed within a closed space in the machine. All tubes in the centrifuge are capped
when spinning and handled (opened) in the tissue culture hood.
Shipping
Have you shipped/will you ship any biological materials related to this study?
Describe the materials you will or expect to ship:
Requested plasmid, vector and virus particles.
Destination (Institution and/or Country):
Various locations
International:
Domestic:
Have you confirmed that it is legal to export each of these materials?
Have you determined what permit(s) will be needed to export?
Have you used/will you use dry ice to pack the materials for shipping?
Who is the primary designated shipper in your laboratory?
Deyan Tong
Yes
Have you received/will you receive a shipment of any biological materials related to this study?
Describe the materials to be received:
Requested plasmid, vectors and cell lines.
Origin (Institution and/or Country):
Various locations
International:
Domestic:
Have you determined what permit(s) will be needed to import each of these materials?
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Applicant comments:
When the materials are requested at that time we will determine if permits are needed to import or export the material. All
shipping will be done by lab personnel with certified training in shipping procedures.
We may ship, or have shipped to us, biological materials related to recombinant DNA, human/mouse cell lines, and/or
replication deficient virus. We anticipate that our lab will be involved in a number of different collaborative relationships
where we will prepare competent virus for infection of human or mouse cell lines; in most of these cases, transfer will occur
between our lab and the neighboring labs at UCSF. Such shipments would be performed in accordance with the UCSF
Infectious Agents Transport Guidelines. Furthermore, all lab personnel responsible for shipping any biohazardous materials
will attend "Safe Shipping of Biohazards" training.
Health Surveillance
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11
Indicate which UCSF health surveillance/immunization programs and critical elements will be placed in effect.
Bloodborne Pathogens Yes
Q-Fever No
Orthopoxiviruses (vaccinia and others) No
Toxins No
Herpesvirus simiae (Monkey B or Herpes B virus) No
M. tuberculosis No
Custom health surveillance/immunization program will be in effect No
Contacts & Personnel
Name
McManus, Michael T.
Degree
Ph.D.
Tong, Deyan
Master
Title
Assistant
Professor
New Academic
Phone
415-502-2049
415-476-4558
Roles
Bio Lab Supervisor, Principal
Investigator
Bio Lab Work, Bio Shipping
Email
mmcmanus@dia
betes.ucsf.edu
Deyan.Tong@uc
sf.edu
Personal Protection Equipment
uniforms, lab coat, disposable outers: Yes
head covers: No
face masks: No
safety glasses: Yes
shoe covers or boots: No
rubber or disposable gloves: Yes
respirator No
other No
Describe procedures for which respirators or other special PPE will be used here:
Cell culture, cell transfection and transduction, biochemistry, cell function assays, flow cytometry. Safety glasses will be used
when operating UV box or handling cryovials from liquid nitrogen vessels.
McManus, Michael T. - Admin/Functional Role:
Laboratory Supervisor :
General Laboratory Work :
Animal Research :
Shipping (to/from campus) :
Local Transportation of Materials :
Billing Contact :
Yes
No
No
No
No
No
McManus, Michael T. - Materials Handled or Exposed To:
Bloodborne Pathogens : Yes
McManus, Michael T. - Training Records
Course Title
Completed Date
Biosafety
11/20/2013
Biosafety Training (was BSL Level II)
09/19/2010
Fume Hood Safety Training (Online)
04/24/2008
Lab Safety Refresher w/Fume Hood 2008
03/24/2009
Laboratory Safety for Researchers
10/25/2005
Laboratory Safety for Researchers (2013)
08/22/2013
UCSF OEH&S Bloodborne Pathogens (Online)
01/21/2015
BU041135-Y03-V21-2015-03-19-Final.doc
12
Tong, Deyan - Admin/Functional Role:
Laboratory Supervisor :
General Laboratory Work :
Animal Research :
Shipping (to/from campus) :
Local Transportation of Materials :
Billing Contact :
No
Yes
No
Yes
No
No
Tong, Deyan - Materials Handled or Exposed To:
Bloodborne Pathogens : Yes
Tong, Deyan - Training Records
Course Title
Biosafety
Lab Safety for Researchers
Safe Shipper - Biohazards (Online)
UCSF OEH&S Bloodborne Pathogens (Online)
Completed Date
12/29/2014
01/07/2015
01/07/2015
12/29/2014
Locations
Parnassus/Health Sciences West [ 1032 ] - Location Detail
Campus: Parnassus
Building: Health Sciences West
Room: 1032
Biosafety Level (BSL): 2
Animal Biosafety Level (ABSL): N/A
Room Use: Procedures, Tissue Culture
Parnassus/Medical Sciences [ 1126 ] - Location Detail
Campus: Parnassus
Building: Medical Sciences
Room: 1126
Biosafety Level (BSL): 2
Animal Biosafety Level (ABSL): N/A
Room Use: Procedures, Storage, Tissue Culture
Equipment
Equipment
BS Cabinet
BS Cabinet
Make
Forma Scientific
LabConco
Model Number
1136
36208 Delta S
Serial Number
13438-201
061163120AD
Location
Health Sciences West/10/1032
Health Sciences West/11/1126
Other safety equipment(s) (e.g. Glove box, etc.):
Certification
I attest that the information contained in this application is accurate and complete. Yes
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13
I agree to comply with the requirements pertaining to shipment and transfer of infectious agents and
materials covered under the Cal/OSHA Bloodborne Pathogens Standard and/or Recombinant DNA.
I hereby adopt the CDC/NIH Biosafety in Microbiological and Biomedical Laboratory, current edition,
and/or the NIH Guidelines for Research Involving Recombinant DNA Molecules, current edition, and the
UCSF Biosafety Manual as the principal biosafety manuals for my laboratory, or, I will provide a
supplemental Biosafety Manual in addition to, or in place of, these references as I deemed necessary or
when specifically requested by the IBC (Institutional Biosafety Committee). I understand that a
supplemental Biosafety Manual must be approved by the IBC for the specific research conducted and
before research can commence.
I recognize that I have responsibility for ensuring that ALL persons who enter my laboratory practice
appropriate biosafety precautions.
I ensure that all personnel listed in this application have received or will receive appropriate and required
training in safe laboratory practices and the procedures for this protocol before any work begins on this
project and at least annually thereafter.
I will follow the health surveillance practices as approved for this protocol and inform ALL persons
working on the protocol, or in shared space, about appropriate emergency assistance information for
their location(s).
I will inform my supervisor, Occupational Health Services at 885-7850 and the Biosafety Officer at 5142824 of any significant research-related accident or illness as soon as possible after its occurrence.
I will submit a request for approval from the IBC of modifications to the study, facilities or procedures.
I will adhere to the UCSF biosafety guidelines referred to in this application.
Shipment of Biological Materials: I will follow University Policy and all applicable Federal and
international regulations whenever I ship biological materials domestically and internationally. I will also
obtain the proper importation or exportation permits / licenses through the Biosafety Office (514-2824)
before shipping to or receiving from any international location any biological material.
BU041135-Y03-V21-2015-03-19-Final.doc
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
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