Genome editing

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Genome editing

I-Stem’s CRISPR platform is dedicated to the production of genetically modified human pluripotent stem cell lines for the study and treatment of single gene disorders.

Since its discovery, the CRISPR-Cas9 genome editing technique has continued to evolve and offer new applications. On the platform, we use the CRISPR-Cas system (CASS) to modify the genome of human induced pluripotent stem cell lines (IPSCs) by indels creation (nucleotide insertion/deletions), base editing or homologous directed recombination (HDR) approaches.

The goal is to generate :

IPSCs with heterozygous or homozygous loss of gene expression
IPSCs “tool” lines to follow the expression of a gene or to induce its overexpression
IPSCs lines carrying genetic mutations detected in a patient for whom no cell sample is available
isogenic IPSCs lines that are controls of patient IPSCs lines in which the genetic mutation has been corrected.

The genetically modified IPSC clones produced by the platform are delivered after ON-TARGETS and OFF-TARGET tests of the CCAS system and quality control (absence of mycoplasma, maintenance of pluripotency and genomic integrity).

Team members

Sandrine Baghdoyan, PhD-HDR

Platform Manager - INSERM

Pascal Fragner, PhD

Platform Engineer CECS

Michel Cailleret

Engineer INSERM

Publications

Pathological modeling of glycogen storage disease type III with CRISPR/Cas9 edited human pluripotent stem cells.

01 January 2023

Rossiaud Lucille,

Fragner Pascal,

Barbon Elena,

Gardin Antoine,

Benabides Manon,

Pellier Emilie,

Cosette Jérémie,

El Kassar Lina,

Giraud-Triboult Karine,

Nissan Xavier,

Ronzitti Giuseppe,

Hoch Lucile,

Frontiers in cell and developmental biology

SEE ALL THE PUBLICATIONS

Team equipment

ThermoFisher Scientifc

Electroporation system Neon

Editing of IPS lines.

This model has a tip chamber that generates a uniform electric field that greatly increases transfection efficiency and cell viability.

SARTORIUS

CellCelector

For high-throughput cloning of edited cells, we use this automatic sampling machine for single cells and colonies.

Agilent

Bioanalyzer Agilent 2100

For evaluation of clone editing, we use this automated electrophoresis system.

Miltenyi - Cellomics ThermoFisher Scentific

FACS & CellInsigth CX7

For the control of pluripotency maintenance, the platform uses a FACS (Miltenyi) or the automated microscope CellInsigth CX7 (Cellomics ThermoFisher Scentific).