Genetic Modified Organisms and containment.
Working with genetic modified organisms needs to be performed according to the GMO-decree and the
GMO-regulations. Before starting handlings, you need to have a permit ., GMO permits are based on the
risk analysis according to attachment 5 of the Regulations.
When you want to perform handlings with gmo’s outside the allowed restriction aria or the approved
restriction level, you have to meet some requirements:
a: These handlings have to be part of the permit too (e.g.microscopy of ML-II cells in a ML-I room, or
microscopy of ML-I cells in a non-gmo laboratory;. FACS analyse of living ML-I or ML-II classified cells at
a FACS that is in a lower classified area. X-ray or Radiation of cells in the Radio Nuclide Laboratory,
classified as D-I has to go to the permit as well.).
b. Open handlings with ML-II organisms are in general only allowed in a Biosafety Cabinet, to prevent the
escape of hazards (aerosols with gmo’s).
Exceptions:
- Lentiviral transduced cells of 2nd and 3rd generation with a SIN transfervector, and a reduction
factor >100. These cells can be downscaled to ML-I, which means that you are also allowed to
perform open handlings at a laboratory bench, since the risk of virus formation is defined as
negelictible. This is explained in a COGEM letter.[COGEM = Committee Genetic Modification,
advisory board of the GMO-office and the Ministry of Infrastructure and Environment]
So far, the GMO office, following the COGEM advises will only allow handlings with 3rd generation
lentiviral transduced or infected cells meeting the above mentioned restrictions when you want to
perform open handlings totally outside containment levels.The risc analysis for the 2nd generation
lentivirus transductions is not performed by the COGEM yet..
-Retroviral transduced cells can only be downscaled to ML-I when these cells meet the reduction
factor >100 and are tested negative with a validated test on PERT (Reversed Transcriptase). This
can be performed at the lab of Joerg Schupbach (University Zurich). The PERT assay detects
retrovirus particles by means of their reverse transcriptase activity. A negative PERT result
means absence of replication-competent RV particles in the investigated samples, while a
positive result needs further investigations as RT activity may also be present in RV particles
not competent of replication.
In order to perform the test a 1 ml cell culture supernatant is needed, precleared in order to
remove cells & cellular debris, frozen at -70°C. Transport on dry ice. This test is accredited,
also by Dutch the GMO office. For non-commercial projects, the costs are €100 per sample.
For information: Diny van Faassen (tel. 81237)or Marjanne Markerink (tel. 82986); Biosafety Officers