1. Home
2. Animal research
3. Ethics & welfare

A multi-site approach to implementing the 3Rs in policy and practice

European Directive 20120/063 EU and its French transposition

PHENOMIN nodes work under European Directive 20120/063 EU and its French transposition (European Directive 2010/063 EU, French « Decree n° 2013-118 » about protection of animals used for scientific purposes):

• Animals we work with are bred in our facilities or come from certified breeders.
• Animal facilities: our institutes have official licenses delivered by the official regional vet (DDPP) to take care of animals, breed them and perform protocols. Housing conditions are standardized and match with the regulation to respect animal welfare.
• Staff: all technicians taking care of mice are specially trained.

Ethical concepts

Following the European Directive also means that we work following ethical concepts:

In 1959, Russel & Burtch have developed the famous 3 Rs concept in order to protect animals used for scientific purposes:

• Replacement: methods which avoid or replace the use of animals.
• Reduction: methods which minimize the number of animals used per experiment.
• Refinement: methods which minimize suffering and improve animal welfare.

Harm-Benefit Analysis (HBA) : the main goal of this analysis is to make a balance between

• The benefit we expect from an experiment like the scientific outcome that can benefit to Humans, Animals or the Environment
• The harm: animal constraining, side effect for example.

In order to replace, PHENOMIN:

• Brings an assistance and awareness to choice and/or design the best model for a research project.
• Makes sure that the use of other in vitro models (molecular, cellular) was made beforehand.

In order to reduce, PHENOMIN:

• Realizes the centralized and coordinated freezing of the murine lineages, and their fast revival in order to limit as possible lineages preservation as respirante shape (living animals).
• Supervises animal cohort generation to avoid reproducing existing models, limiting the production of animals on a national scale and rationalizing their studies in pipeline.
• Develops modern functional analysis which are not invasive or little invasive, allowing a longitudinal follow-up: ultra-sound, photo-acoustic imaging, fluorescence to follow cell lineage (cytometry), telemetry, …( "Experimental rodent cancer models: contribution of preclinical imaging to the 3Rs strategy", STAL- Vol47, p15, 2019 (1.4 MB) ) -
• Uses standardized functional analysis in pipeline to obtain the maximum of scientific parameters and information from the same animal cohort to conclude about a phenotype, and in a standardized way (example: www.mousephenotype.org).
• Uses bio-statistic analysis, and evaluate the power of the statistical tests realized in order to estimate in the closer the number of animals which are needed to include in the studies.

We are involved in the setting up of an alternative to reduce the number of animals used and improve the quality control to determine the ability of a mutant mouse line to recover after cryopreservation. Our recent publication (Blastocyst genotyping for quality control of mouse mutant archives: an ethical and economical approach. Transgenic Res. 2015; 24(5): 921–927. F Scavizzi et al.,2015) described a new technical approach which is now routinely used in PHENOMIN projects.

PHENOMIN has implemented electroporation for the delivery of CRISPR/Cas9 with or without small donor DNA. This method allows the drastic reduction of the number of donor females and fertilized oocytes. We consistently obtained high efficiency mouse genome editing for multiple genes, and we successfully generated mouse embryos with a variety of editing schemes, including indel mutations, point mutations, genomic deletions, and small precise insertions. Taken together, CRISPR-electroporation is a simple, economic, 3Rs friendly, high throughput, and highly efficient technique for genome editing in vivo, which can replace the traditional microinjection-dependent genome editing technique in mice and in other mammalian species ( Generating rodent model by CRISPR/Cas9 genome editing (480.9 KB) ).

In order to refine, PHENOMIN:

• Pursues the improvement of the breeding and housing conditions of the rodents with respect to their physiology and socialization behavior.
• Reduces the use of the invasive protocols by deploying to the non-invasive methods as the imaging, the follow-up by telemetry.
• Insures the training, the transfer of the best practices for delimiting the “end point” of the studies.
• Defines pipeline of standardized preclinical analysis (for example: studies in cardiology, immunology, then neurology on the same animal cohort) to maximize the kind and amount of information collected within the same project from a limited animal number.

Ethics committees and project authorization

All PHENOMIN projects that involve animals have received an approval from our ethic committees and an authorization from the Ministry of Research.

Projects evaluations are realized following the 3Rs and HBA concepts with a huge attention on refinement (use of anesthesia, of painkillers when needed, respect of good practices).

Following the national convention that our ethics committees have signed, our members have different point of view being researchers, technicians, vets or "naive" representing the community. Moreover, our teams are trained to perform those evaluations properly (the GIRCOR has set up trainings dedicated to ethical committee members). 

Animal Welfare Bodies

Projects authorizations are released before starting. Once they have started, they are followed on the field by our Animal Welfare Bodies (AWB).

In addition to this mission, AWB main missions are advising research teams, answering ethical and animal welfare questions, visiting animal facilities to check the well going of protocols and developing an ethical way of working with animals in the long term. Thus, AWB are proactive to improve animals’ living conditions.