platforms 78 platforms 79 Viral vectors platform In 2023, the viral vectors platform strengthened partnerships with E-Zyvec/ Polyplus/Sartorius for plasmid design/synthesis and CER groupe for upscaling AAV and Lentiviral vector production. The platform evaluated a new helper virus to enhance AAV production and quality, securing a Win4Excellence grant (GT4Health) for advanced vector production strategies. In 2024, the platform plans to develop new packaging cell lines to boost AAV production and create AAV coupled with nanobodies for better targeting. Additionally, the platform will offer new protein production services (in mammalian cell lines) in collaboration with the ULiege Centre for Protein Engineering (CIP). Gene Editing platform The platform, established in 2022, supports projects by creating genetically modified mouse models and performing precision edits into cell lines. Our current portfolio ranges from NHEJ-based knock-out to simple knock-in alleles (SNP replacement, tag insertion). The platform has also expertise in the generation of CRISPR modified zebrafish. Because delivery of CRISPR tool is sometimes a barrier into some type of cells, the approaches are versatile and use preferentially RNPS but also lentiviral viral vectors (in close collaboration with the Viral Vectors platform). In 2023, the platform expanded to editing cancer cell lines and producing KO mice, exploring new editing strategies beyond CRISPR-Cas9. These tools facilitate gene function research and genomic validation. Future plans include developing pipelines for precise genetic mutant cell lines using Prime Editing and employing AAV vectors for extensive genome modifications in mice. In 2023 Emmanuel Di Valentin, GIGA-Viral Vectors Manager Hélène Pendeville, GIGA-Gene Editing Manager edivalentin@uliege.be hpendeville@uliege.be www.gigaviralvectors.uliege.be www.gigageneediting.uliege.be Contact Services VIRAL VECTORS Tailored made design of plasmid for viral vectors production in collaboration with suppliers such as e-Zyvec/polyplus and VectorBuilder Production of custom viral vectors AAV, retro and lentiviral vectors Cell lines modifications using lentiviral vectors Gene overexpression Tagging Protein depletion (siRNA/hRNA) KO/KI (using CRISPR technology) Cell immortalization GENE EDITING Know how A la carte solutions for cell lines, zebrafish or mouse CRISPR projects In silico design of guide(s) RNA and donor constructs Vectorization of sgRNA and Cas in viral vectors (adenovirus, lentivirus) Services in cell lines Cas9 expressing cell lines Generation of CRISPR KO/ CRISPR KI pools Illumina sequencing and analysis of CRISPR KO/CRISPR KI pools Expansion and screening (by PCR or NGS) of clonal populations CRISPRa/ CRISPRi pools (with viral vectors) Services in mouse and zebrafish Design of the project (KO, KI, deletion, small tags) Microinjection/ electroporation of CRISPR reagents in zygotes Embryo transfer in pseudopregnant females (mouse) Blastocyst (mouse) or embryos (zebrafish) genotyping Screening of F0 animals Management of the F0 founders and screening of the F1 heterozygous animals x
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