EdGe | Genome Editing

Anthony Gobert is managing the technical platform “Genome editing” (EdGe) of the IBMP and proposes to guide you in your projects for the production of targeted mutants (KO or others).

CrispR systems are becoming more efficient and are constantly evolving. The target of the Cas enzymes is PAM (protospacer adjacent motif) dependent (e.g. NGG for SpCas9). To cleave targets throughout the genome, a range of Cas enzymes with different PAMs was cloned into an in-house cloning-transformation system (see table).

The EdGe technical platform also offers solutions for targeting several areas on the genome or multigenic families with vectors that can contain two guides or allow the multiplexing of guides (see table).

Available : a couple of vectors for prime Editing, an ultra-precise genome editing solution based on a sequence present on a modified RNA guide (pegRNA) called ‘primer’ containing the desired mutation, a tag, or any other sequence. This system is not based on homologous recombination that is ineffective in most plants but on the reverse transcription of the “primer” (see table).

Technological watch: the EdGe technical platform seeks to acquire the most efficient systems published to date and to introduce them into the in-house destination vector for an ever faster production of mutants.
Protocols are available as well as my increasingly precise advices thanks to the feedback from users of the technical platform.