Custom Cell line development using HAC/MAC vector in various host cells
Using our technology, we can introduce a very large gene or multiple genes and create stable cell lines. The genes will be very stably expressed, and cells can be stably passaged many generations without integration into the host genome. Once the human artificial chromosome construct is created, it can be easily introduced into various recipient cell lines. By introducing the HAC/ MAC vector to ES cells, we can also make transgenic animals.
- Applications include:
- Creation of stable cell lines for drug screening
- Creation of stable cell disease models from various iPSC lines
- Evaluation of drug effects on multiple genes
- Expression of difficult to express genes
- Expression and production of large protein complexes
- Creation of ES cell lines, and then creation of in vivo model
Example of gene expression using HAC/MSC vector
| Introduced gene | DNA type | Number of genes/size | Recipient cell | Description |
| Protein expression | ||||
| CMV-human EPO | cDNA | 1 gene | HFL-1 | Construction of erythropoietin expression system |
| CAG-human FVIII | cDNA | 1 gene (16 copies)(200kb) | CHO hprt-/-, hiMSC | Expression of FVII (16 copies) |
| HSP70-insulin | cDNA | 1gene | HT1080 | Expression of insulin from HSP promoter |
| Gene therapy | ||||
| Human dystrophin | genomic | 1gene (2.4 Mb) | hiMSC, mouse, mdx-iPS, DMD-iPS | introduction of HAC vector containing dystrophin gene into iPS cells |
| Mouse CD40L | Genomic | 1gene (12 kb) | Jurkat, U937 | Expression of CD40 ligand |
| Human P53 | Genomic | 1gene (150 kb) | mGS p53-/- | Introduction of hP53 gene into endogenous P53 gene-deficient mGS cells |
| Gene therapy system | ||||
| TR-DNA-PKcs | cDNA | 1gene | V3 | Control of DNA-PKcs expression by Tet system |
| Ubc-hTERT-IRES-GFP | cDNA | 2 genes (IRES-mediated Polycystronic expression) | HFL-1 | Prolongation of life of normal fibroblast by expression of telomerase reverse transcriptase |
| OPN-EGFP | cDNA | 1 gene | hiMSC | Lineage specific gene expression by OPN promoter |
| Screening system | ||||
| OC-GFP | cDNA | 1gene | CHO | Screening system for substances with osteogenic activity |
| Others | ||||
| Human CYP3A cluster | Genome | 4genes (0.7Mb) | Mouse (in vivo) | Creation of mouse model expressing human metabolizing enzymes |
| Yamanaka four factors+ p53ph RNA | cDNA | 5 genes (11 copies total) (40kb) | MEF, mouse iPS | iPS cell generation |
