Skip to main content

Comet assay and cell array for fast and efficient genotoxicity testing

Objective

A battery of reliable and validated in vitro assays is needed to test for genotoxic and cytotoxic effects of chemicals (as in the REACH programme) without resorting to animal experiments. The comet assay, a sensitive indicator of DNA damage, will be combined in this project with the Cell Array system, to establish and validate high capacity assays suitable for chemical testing. Up to 800 cell samples will be processed for comets on a single microscope slide. Arrays will use cells with different metabolic capabilities, and data on cytotoxicity will be obtained in parallel with DNA damage. A medium-throughput assay will also be developed. Comet analysis by differential staining of damaged/undamaged DNA using established and novel dyes combined with automated image analysis will be faster and more reliable than at present.
A crucial aspect of the cellular response to DNA damage is DNA repair; variations between people can affect cancer risk. Genotoxic chemicals can act by interfering with repair.
Two methods for measuring repair, one based on the comet assay, the other using a 'Repair Chip' approach, will be compared. In addition, fluorescent probes which lock onto the DNA after hybridization ('padlock probes') will be applied to study gene-specific DNA repair. The modified comet assay methods will be assessed for reproducibility and sensitivity in an inter-laboratory prevalidation trial, using a coded set of standard chemicals, to satisfy regulatory bodies as well as industrial users of the technology. The result will be robust, validated, high-throughput in vitro genotoxicity tests. This proposal brings together academic partners and SMEs, together with a large industrial concern, and the European Centre for the Validation of Alternative Methods (ECVAM). Dissemination through professional and trade publications, regulatory channels, and scientific conferences will lead to widespread adoption of the new methods.

Field of science

  • /natural sciences/physical sciences/astronomy/planetary science/comets
  • /natural sciences/biological sciences/genetics and heredity/dna

Call for proposal

FP6-2005-LIFESCIHEALTH-7
See other projects for this call

Funding Scheme

STREP - Specific Targeted Research Project

Coordinator

UNIVERSITETET I OSLO
Address
Problemveien 7
Oslo
Norway

Participants (14)

IMSTAR S.A.
France
Address
60 Rue Notre Dame Des Champs
Paris
JOINT RESEARCH CENTRE
Belgium
Address
Rue De La Loi 200
Brussels
NORWEGIAN INSTITUTE OF PUBLIC HEALTH
Norway
Address
Geitmyrsveien 75,0462 Oslo 4
Oslo
SLOVAK MEDICAL UNIVERSITY
Slovakia
Address
Limbova 12
Bratislava
CHIRON AS
Norway
Address
Stiklestadveien 1
Trondheim
TATAA BIOCENTER AB
Sweden
Address
Medicinaregatan 8A
Göteborg
ASTRAZENECA UK LIMITED
United Kingdom
Address
Stanhope Gate 15
London
THISTLE SCIENTIFIC LIMITED
United Kingdom
Address
71 Ochiltree
Dunblane, Perthshire
SEVERN BIOTECH LTD
United Kingdom
Address
Park Lane, Kidderminster
Kidderminster, Worcestershire
BIOPREDIC INTERNATIONAL
France
Address
8-18 Rue Jean Pecker
Rennes
UPPSALA UNIVERSITY
Sweden
Address
Sankt Olofsgatan 10B
Uppsala
COMMISSARIAT À L' ENERGIE ATOMIQUE
France
Address
Batiment Le Ponant D - 25 Rue Leblanc
Paris
INSTITUT NATIONAL DE LA SANTÉ ET DE LA RECHERCHE MÉDICALE
France
Address
101 Rue De Tolbiac
Paris
USTAV EXPERIMENTALNI BOTANIKY - AKADEMIE VED CESKE REPUBLIKY
Czechia
Address
Rozvojová 135
Praha 6 - Lysolaje