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StemEdit Clinical Gene Editing Service

Thanks to the discovery of CRISPR-Cas9, gene editing is more accessible than ever before. However, some genetic modifications remain challenging and there are even more factors to consider if your cells are intended for clinical use.

By outsourcing your clinical gene editing to REPROCELL, you can achieve the genotype you need before moving on to expensive cell bank manufacturing processes. Our StemEdit service uses advanced CRISPR-SNIPER* gene editing technology to develop your engineered stem cells. Due to the increased screening specificity of CRISPR-SNIPER, we can successfully achieve complex genetic edits with high accuracy at an early stage. By evaluating the percentage of target cells and determining their transfection efficiency (go/no-go decision point) StemEdit saves time as the SNIPER pre-screen is performed before laborious clone selection.

REPROCELL Stemgent™ – the stem cell experts

Case study: HLA Disruption in clinical Seed Stock iPSCs

One limitation of using iPSCs clinically is immunogenicity caused by Human Leucocyte Antigen (HLA) mismatching, which can reduce the in vivo survival and therapeutic efficacy of these cells.1,2 The immunogenicity of iPSCs can be reduced by disrupting the genes responsible for immune recognition using CRISPR-Cas9. One approach is to knock out the B2M (β2 microglobulin) and CIITA (major histocompatibility complex [MHC] II transactivator) genes.

In humans, the B2M protein is required for the presentation of HLA protein A-G on the cell surface, while CIITA is essential for HLA-II transcription.2,3 Dual knock-out of the B2M and CIITA genes disrupts the presentation of MHC-I/MHC-II proteins in iPSCs, improving their therapeutic potential while maintaining pluripotency.1 At REPROCELL, we offer this knock-out as a service; the diagram below shows our workflow for the single knock-out of B2M and CIITA genes, with phase contrast images of the Seed-Stock iPSCs at each stage. 

 

iPSCsiPSC
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DNA manipulation plastic modelGene editing B2M KO
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B2M KO iPSCsB2M KO iPSCs
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DNA manipulation plastic modelGene editing CIITA KO
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B2M CIITA knockout iPSCs 2B2M/CIITA KO iPSCs

Interested in finding out more about StemEdit?

If you are interested in finding out more about REPROCELL's StemEdit Clinical Gene Editing Service, please click on the button below to make an inquiry. Our stem cell experts will be happy to arrange a free consultation to discuss your project in more detail. For further information about our clinical iPSC services, please refer to our Clinical iPSC FAQ page. 

References

1. Wang X et al. Diminished expression of major histocompatibility complex facilitates the use of human induced pluripotent stem cells in monkey. Stem Cell Research & Therapy 11:334 (2020).

2. Xu H et al. Targeted Disruption of HLA Genes via CRISPR-Cas9 Generates iPSCs with Enhanced Immune Compatibility. Cell Stem Cell 24 pp566-578 (2019).

3. Deuse et al. Hypoimmunogenic derivatives of induced pluripotent stem cells evade immune rejection in fully immunocompetent allogeneic recipients. Nature Biotechnology 37 pp 252–258 (2019).

*SNIPER = Specification of Newly Integrated Position and Exclusion of Random-integration. Note: CRISPR-SNIPER modified cells are developed, manufactured or supplied by GenAhead Bio Inc. under license from ERS Genomics Limited.