University of Sheffield
Local Genetic Modification Safety Committee
THE GENETICALLY MODIFIED ORGANISMS (CONTAINED USE) REGULATIONS 2000
FURTHER DETAILS
http://www.hse.gov.uk/dst/acgm27.htm
http://www.hse.gov.uk/hthdir/noframes/acgmcomp/acgmcomp.htm
RISK ASSESSMENT FORM (REV6 01/2002)
This form should be used for preliminary assessment of all GMM proposals and for local approval
of Class 1 activities. Note that use of the “Brenner Scheme” is now discouraged.
If you believe aspects of your work should not be disclosed please complete your forms with
reference to: http://www.hse.gov.uk/dst/confid.htm.
Provide sufficient information for each table entry to justify your proposal.
Proposer:
Status (eg staff/PG): Research Staff
Head of research group
(This person is responsible for
project):
Dr. Marcelo N. RIVOLTA
Address of research laboratory
Rm C08
Biomedical Sciences
Quad Building
Western Bank
S10 2TN
Telephone: 0114 222 222385
Fax: 0114 2222360
E-mail:
m.n.rivolta@sheffield.ac.uk
Title of project: Manipulation of mouse inner ear stem cells
Overview or aim of the project:
The aim is to use conditionally immortal mouse cell lines to model developmental events in the
differentiation of sensory hair cells, neurons and epithelial cells in the inner ear. Cells are cultured in
vitro and transfected to study gain and loss of function for selected genes. They are also co-cultured
with similar cell lines and organotypic primary cultures from the mouse. Cells were immortalised
with a stably-integrated, temperature-sensitive variant of the T antigen from the SV40 virus. They
were derived from a transgenic mouse (Jat et al. PNAS, 88:5096-5100, 1991).
List all workers (staff and students)
involved in the work, their status
(eg PG, lecturer, technician,
postdoc) and include date of GMM
medical if known
Name: M. N.RIVOLTA
Status: Senior Research Fellow
Date of Medical: Bristol 1997
Room and building where work will be carried out: Quad Building C01
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Hazard identification in respect of human health & environment
(http://www.hse.gov.uk/dst/acgm27a.htm, http://www.hse.gov.uk/dst/acgm28b.htm#1
& http://www.hse.gov.uk/hthdir/noframes/acgmcomp/2a.pdf). Include:

Hazards associated with the
Host (note that separate
assessments are required for
each distinct host/vector
system)
The conditionally immortal cell lines are disabled, noncolonising and non-pathogenic to humans or animals. They
have limited survivability in the environment and culture
requirements unlikely to be satisfied outside of the laboratory.

Hazards associated with the
Vector
The vectors are unable to infect or transfer DNA to other hosts.
All are puc18 or pbr322 derived plasmid vectors.

Hazards arising directly
from the insert (origin,
suspected function)
The transgene is stably integrated into the cells, which were
derived from a transgenic animal. Immortalisation is via a
temperature-sensitive variant of the A gene from SV40.

Hazards arising from the
alteration of existing
pathogenic traits
No detectable change in, or history of, pathogenicity.

The potential hazards of
sequences within the GMM
being transferred to related
organisms.
The potential hazards are negligable and cells immortalised in
the same manner have been used widely for 20 years.

Consideration of the
likelihood that, in the event
of exposure, the Final
GMM could actually harm
human health
No significant hazards identified and it is unnecessary to
consider predicted properties with respect to human health
issues. There is a long history of safe use.
Hazard Group 1
2
Estimate severity or consequences of any harmful effect were it to occur. Provisional Class and
containment level. Include:
Hazard group 1. No significant hazards identified.
 Hazard group of organism

Other pathogens
Containment Level 1.

Will modification affect
basic hazard classification?
No
Environment and activity considerations (http://www.hse.gov.uk/dst/acgm27e.htm). Include:
Low.
 Likelihood of hazards

Scale of work

Control measures and waste disposal
routes to be used to protect the
environment
(http://www.hse.gov.uk/hthdir/noframes/acg
mcomp/3a3.pdf &
http://www.hse.gov.uk/dst/acgm29c.htm#4)

Methods used to validate inactivation
of waste prior to leaving premises (eg
source of chemical kill data and its
applicability to hazard and associated
organic elements or autoclave testing
programme)
(http://www.hse.gov.uk/dst/acgm29c.htm#2)

Revision of containment

Check that hazards controlled by
containment
Assign final activity Class. Compare
containment and control measures to control
the risk as detailed in Schedule 8
(http://www.hse.gov.uk/dst/acgm27g.htm).
All non-Class 1 activities must be notified on
the appropriate form
(http://www.hse.gov.uk/dst/conduct.doc).
Small scale laboratory work. Maximum of 200 ml in
tissue culture flasks.
Cells are cultured in a designated room and
manipulated in a Class 2 hood. Spillages are easily
treated with alcohol and disinfectant. For laboratory
operations a standard containment level 1 facility
with good cell culture practice will be sufficient to
limit contact with humans and the environment.
All waste is incinerated. These measures are
primarily designed to prevent contamination and are
in excess of what would be required for protection of
human health or the environment. If containment
was breached the risk would effectively be zero.
We use yellow bins provided by a licensed waste
disposal company.
The bins are validated for disposal of all waste.
Class 1
Date of proposal
The Group head is responsible for ensuring that all workers involved in this project are
proficient in the proposed techniques or ensuring that inexperienced workers will be
supervised throughout training.
Signed by
Proposer:
Group head:
Head of Department:
Departmental BSO
INFORMATION SOURCES
Search the HSE site for GMM regulations and ACGM newsletters
http://www.hse.gov.uk/hsehome.htm
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APPROVAL
This project has been considered and approved by the Local Genetic Modification Safety
Committee of which I am the chairman.
Signed by ……………………………………… Date …………….……..
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