PHENOMIN, as a large scale national infrastructure for Biotechnology and Health, provides services for all French laboratories.
PHENOMIN offers 37 different services in genetic engineering, model validation experiments, state-of-the-art zootechnic and phenotypic approaches, archiving and distribution. These services benefit to users who conduct genetic researches combining generation, validation and phenotyping analysis of mutant mice model (physiological, behavioral and anatomical analysis).
CRISPR Mediated REarrangement (CRISMERE) strategy
By adapting CRISPR / Cas9 technology, researchers of PHENOMIN-ICS have been able to quickly and efficiently generate mouse models of diseases related to variations in the number of copies of certain regions of the genome. These results are published in Scientific reports (March 2017).
- The CRISPR / Cas9 "revolution"
Discovered in 2012, the technique known as "CRISPR / Cas9" allows to target DNA very precisely through a guide RNA. The latter is associated with an enzyme, Cas9, which makes it possible to cut, replace, inactivate or modify the targeted gene. More precise, faster and cheaper than all the techniques used to date, this method is now used in many laboratories to modify cells in vitro. Numerous studies have shown that its application in vivo is possible but the effectiveness of the methods remains limited in these cases.
- Easily create mouse and rat models
To date, to create murine models, researchers use methods combining the Cre-Lox recombination system and the use of embryonic stem cells, but these processes are long and tedious, and it takes about 3-4 years to create a new mouse model. Here, scientists have adapted the CRISPR / Cas9 technique for the creation of mutant animals. They have doubled the RNA-guides, using two very similar guides to increase the efficiency of the break and have thus managed to modify important regions of the DNA (up to 24.4 Mb). Deletion, duplication, inversion, many configurations have been tested and validated by the generation of mouse lines carrying these mutations, and their transmission to the lineage. "The size of the targeted DNA section is not really a limiting factor, it's more the question of viability due to the removing certain genes that limits us" explains Marie-Christine Birling. Conclusion: it is now simple and fast (6-7 months) to create new disease models, good news for translational research.
- Mice and rat models of trisomy 21
With this new method developed at the PHENOMIN-ICS, Yann Hérault and his team at IGBMC have created new mouse models of trisomy 21, a disease due to the presence of an additional copy of the chromosome 21.
This was not possible with the previous techniques, but the researchers also succeeded in performing the same manipulations in the rat, thus creating an interesting model for evaluating behavior, knowledge, memory and social interactions. According to them this technique is adaptable to any species and its use could be made on human cells.
This study has been financed by the ANR, the Fondation Jérôme Lejeune and the Fondation Bettencourt-Schueller.
Charles River Europe and Phenomin-ICS Launch New Collaboration for Model Creation
This exciting partnership offers our European clients a complete integrated solution for mouse and rat models, from creation to validation. Our combined in vitro and in vivo expertise shapes a broad portfolio of design (ES cell, CRISPR/Cas9, random DNA insertion), breeding and associated services to create and maintain the most relevant models for your studies.Rely our industry-leading germ line transmission, strict quality controls, fast turnaround and dedicated project managers to help you successfully reach your goals.
Genetic Engineering Design Options
CRISPR-Cas9 is a rapid and cost effective technology to generate mouse and rat mutant models. Now we can offer the generation of knock-out models though direct injection into fertilized mouse oocytes of CRISPR sgRNA and Cas9 mRNA. This approach allows to obtain models in shorter timelines and with reduced costs compared to gene targeting by homologous recombination in ES cells.
Our offer involves:
- Design of an optimized strategy including prediction of off-target sites
- Preparation of sgRNA and Cas9 mRNA
- Injection of sgRNA and Cas9 mRNA into zygotes, to generate F0 mutated animals
- Genotyping of F0 animals and delivery of F0 candidate
- Optional steps:
- Off-target analysis in F0 founder mice (optional)
- Germline breeding of F0 founder mouse to achieve germ line transmission (optional)
- Off-target analysis in germline transmitted F1 mice (optional)
The rat model offers better clinical characterization approaches than mouse model in some cases.
Because rat shares many physiological similarities with humans, the rat has long been a model favored by physiologists, pharmacologists and neuroscientists.
Their larger size allows to perform several experiments, surgical procedures and to monitor physiological parameters that are difficult or impossible in mice. Rat is a key model for cardiovascular disease studies: atherosclerosis, hypertension, stroke models, not only because of its size, but also because of its physiology. By their ability to learn and accomplish sophisticated behavioral tasks, rats are relevant model for nervous system diseases.
Now, CRISPR/Cas9 genome editing opens a new area in rat genetics.
- 1- Jacob, H.J. & Kwitek, A.E. (2002) Multifactorial genetics: Rat genics: attaching physiology and pharmacology to the genome, Nat. Rev. Gen., 3(1), 33-42.
- 2-Cozzi J. et al., (2008) Use of genetically modified rat models for translational medicine, Drug Discovery Today, 13(11/12), 488-94.
- 3- Aitman TJ et al., (2008) Nat Genet. Progress and prospects in rat genetics: a community view. 2008 May; 40(5):516-22.
CRISPR/Cas 9 frequently asked questions
CELPHEDIA WP1 workgroup gathers experts using CRISPR/Cas9 system in various animal models including mouse, rat, zebrafish and drosophila. By sharing our knowledge, experience and an extensive bibliographic review, we have worked together to write some recommendations for CRISPR/cas9 system. Our goal is to help scientists who want to use this technology for gene editing by summarizing current advances on many technical aspects, from the RNA guide optimized design to the genotyping analysis and the validation of the newly generated models.
Archiving and Distribution
- Cryopreservation (Embryos-sperm)
- Distribution (Embryos-sperm, living mice)
Genetic Engineering and Model Validation
- Trangenesis (plasmid, BAC)
- Targeted Genetic Engineering and Model Validation (ES cells)
- Genetic Engineering and Model Validation through IKMC recombinated ES cells
- Production of recombinated ES cells
- Clinical phenotyping
- In vivo imaging
- Drug screening, derisking strategy
- Bio imaging under BSL3 conditions
- In vivo disease models
- Health sanitary controls
- Maintenance of heath status
- Housing and breeding
- Validation of mouse models