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Brightfield microscope image of AST-9650 hiPSCs

ActiCells™ RUO TARGATT™ Hypo hiPSC Knock-in Kit

SKU AST-9650 Categories , , ,

ActiCells™ RUO TARGATT™ Hypo hiPSC Knock-in Kit

Rapidly advance your allogeneic development with a hypoimmunogenic iPSC platform that has pre-built TARGATT™ large knock-in technology.

  • Favorable development profile—reprogrammed from CD34+ cord blood cells for reduced immunogenicity and fewer somatic mutations compared to reprogrammed adult cells 
  • Hypoimmunogenic—B2M/CIITA double knock-out eliminates expression of HLA Class I and II molecules to avoid recognition by CD8+ and CD4+ T cells  
  • Large knock-in ready—quickly add additional immune evasion and disease modifying genes at the reliable H11 safe harbor locus using TARGATT large knock-in technology 
  • Excellent differentiation potential—we’ve successfully differentiated the parental line into a number of cell types, including cardiomyocytes, NK cells, T cells, monocytes/macrophages, and endothelial cells 
  • Commercialization-ready—genome editing performed with Mad7 and TARGATT technology for a single, cost-effective license and clear freedom to operate
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Overview

Reprogrammed from CD34+ cord blood cells and rendered hypoimmunogenic by knocking out both the β2 microglobulin (B2M) gene and the HLA class II transactivator (CIITA), the ActiCells™ RUO Hypo T hiPSC Knock-in Kit (Cat. # AST-9650) is a fast and reliable way to build your allogeneic cell-based medicine.

Designed to jumpstart your allogeneic development, the ActiCells™ RUO TARGATT™ Hypo hiPSC Knock-in Kit gives you a hypoimmunogenic cell line that is immediately ready for further modification. Knock in additional immune evasion elements to avoid clearance by NK cells, macrophages, and dendrites while also expressing disease-modifying genes. The power of the TARGATT™ platform means you can insert all these genes in a single reaction—up to 20 kb—with additional genes in nested reactions. 
The other advantage of TARGATT™ technology lies in its placement at the H11 safe harbor locus, which avoids the potential for silencing and harmful random integration that you get with lentivirus and transposon-mediated integration and consistently delivers gene expression.*
ActiCells™ RUO TARGATT™ Hypo hiPSCs will be the research-matching isogenic cell line to the ActiCells™ GMP TARGATT™ Hypo hiPSCs which are in development.
Don’t need TARGATT™ technology? We also offer ActiCells RUO Hypo hiPSCs (Cat.# ASE-9550) as a cell line only product. 
Each ActiCells™ RUO TARGATT™ Hypo hiPSCs Knock-in Kit contains sufficient reagents for 3 transfections and includes: 
  • AST-9650-C ActiCells™ RUO TARGATT™ Hypo hiPSCs 
  • AST-3091 TARGATT™ 46 CAG-MCS Cloning Plasmid   
  • AST-3084 TARGATT™ CAG-Integrase Plasmid 

Every lot is tested for viability following recovery from cryopreservation, functional pluripotency (formation of the three germ layers), expression of pluripotency markers (OCT4, SOX2, NANOG, SSEA-4, and TRA-1-60), presence of alkaline phosphatase (AP), STR, sterility, and for the absence of mycoplasma and pathogens (CofA available upon request).

Table Header Table Header
Cat. #
ASE-9650
Reprogramming method
Episomal
Donor tissue
CD34+ Umbilical Cord Blood Cells
Gender
Male
Clinical information
Normal
Quantity
1 x 106 cells/vial

*Constructs must have already been shown to be expressable in iPSCs or the differentiated cell type.

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How it works

Cell therapy rejection is, in part, mediated by CD8+ T cells (cytotoxic T cells), which interact with HLA class I molecules, and CD4+ T cells (helper T cells), which interact with HLA class II molecules (Figure 1).
Figure 1. Avoiding the host T cell response is critical for overcoming allograft rejection.

To make hypoimmunogenic hiPSCs, we eliminated cell surface expression of HLA I and HLA II molecules by knocking out two genes (Figure 2):

  • B2M, which encodes the ß2-microglobulin protein, a key part of the HLA class I complex
  • CIITA, the HLA II transactivator which is essential for transcription of HLA class II genes
Figure 2. Knocking out cell surface expression of HLA I and HLA II eliminates the T cell response, although other gene edits are needed to avoid innate immune responses. To eliminate cell surface expression of HLA I and HLA II we created a homozygous double knock-out of the B2M and CIITA genes in a hiPSC line containing a TARGATT landing pad.

Learn more about our approach to developing hypoimmunogenic hiPSCs by watching our webinar, “The Future of Allogeneic Therapy: ASC’s Hypoimmunogenic Donor Cells.” 

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Supporting data

Sanger sequencing chromatograms showing B2m and CIITA knock-out
Flow cytometry data

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