TEFOR-TACGene

TEFOR-TACGene is a facility created in 2011 within Inserm U1154/ CNRS UMR7196, to facilitate access of academic laboratories to genome editing techniques.

Genome editing techniques allow within one cell, the precise modification of targeted DNA sequence in a genome of interest by insertion, deletion or replacement. These changes are based on the DNA repair mechanisms of host cells and the use of specific custom endonucleases that act as molecular scissors. The developments and applications of genome editing with custom endonucleases began in 2005 with Zinc-Finger Nucleases (ZFN), followed in 2011 by the TALE-Ns, for "Transcription-Activated like Effector Nucleases". Since 2013, the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9, which is very easy to implement, makes genome editing accessible to the whole scientific community. The applications of these tools are countless and revolutionize genetics in virtually all living organisms.

Enriched by the rapid evolution of these technologies, TEFOR-TACGene has developed a solid expertise in the design, production and use of TALE-N and CRISPR / Cas9 systems, both for its own research projects in cultured cells and in collaboration, in many model organisms, including the rat, zebrafish, Drosophila, and Xenopus as part of the National Infrastructure in Biology and Health TEFOR supported by Investissement d'Avenir programme (2012-2019). The research and development activities conducted by TEFOR-TACGene aims to provide the community with advanced expertise in tools and strategies of genome editing and develop original solutions for specific issues (such as improving KI efficiency).

Genome Editing Technologies

  • Develop and promote the use of Genome editing techniques using TALE-N and CRISPR / Cas9 system
  • Understanding the mechanisms involved and improve Knock-Out (KO) and Knock-In (KI) efficiencies
  • Maintain an active technology watch in the field

Project Management

  • Offer customized solutions for specific applications of the academic scientific community
  • Propose custom services for the production of vectors, RNA or purified proteins to develop specific applications
  • Test nucleases produced in cell lines or in vitro from purified proteins
  • Analyze the mutations induced by nucleases by genotyping (T7-endonuclease I test or sequencing with PGM)
  • Advise on the use of the technologies and helping research labs to implement them using optimized protocols

CRISPR/Cas9 revolution

INSERM continuous training on revolutionary CRISPR/ Cas9 technology. This training occurs in two phases and will be multisite.

Phase I consists of a critical assessment of CRISPR/Cas9 technology and will occur over 3 days.

Phase II consists of technical sessions. A general practical education will be provided at the National Museum of Natural History in Paris. Trainings to generate genetically modified models in specific organisms will be provided on different sites according the speciality: zebrafish model at AMAGEN (Gif-s-Yvette), rat model at the TRIP (Nantes) and mouse model at Anexplo (Toulouse).

The aim: This workshop will offer an overview of the latest breakthroughs of CRIPSR/Cas9 technology and the challenges of the future, as well as an overview of the various applications.

INSERM training (3 days + practical training): last session in 2016.

Improvement of the Knock-In activity

One of the great challenges in many biological systems is to succeed in improving knock-in (KI) efficiencies to allow better controlled and precise genome modifications. In this context, TEFOR-TACGene seeks to:

  • Develop KI techniques using modified oligonucleotides in cultured cells and in model organisms
  • Understand the molecular mechanisms involved in different cell types and organisms to explain the differences in efficiencies
  • Develop original methods to stimulate KI efficiencies.

Development of different types of nucleases

  • Develop the production of purified proteins (TALE-N, S.Pyogenes Cas9, S.Pyogenes Cas9 nickase, N. Meningitidis Cas9 ...)
  • Test the different constructions and check for their effectiveness

Develop bioinformatics tools for genome editing

  • web site tefor.crispor.net for selection of guide RNA sequences and design of genome editing experiments
  • software for analysis of PGM sequencing of DNA targeted with artificial sequence-specific nucleases

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