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Description
Human iPSC-Derived Neural Stem Cells that have been genetically edited using CRISPR-Cas9 technology to introduce the P301L mutation (CCG>CTG) into the MAPT gene. This line is heterozygous for the P301L mutation where one allele contains the mutation and the other allele is wild type at the locus. Click on the product images to see the data and further details.
The P301L mutation MAPT has been implicated in frontotemporal dementia and parkinsonism (Dumanchin et al., 1998). The mutation affects only the 4R isoforms of MAPT since the exon containing the mutation is spliced out of 3R isoforms (Hutton et al., 1998). Aggregated MAPT/tau protein in affected patients consisted mainly of the 4R isoform (Hutton et al., 1998). The P301L mutation promotes aggregation of MAPT/tau protein into ordered paired helical filaments and beta sheet formation in vitro (von Bergen et al., 2001).
Product Specification
| Starting material | Dermal fibroblast |
| Donor gender | Female |
| Donor age at sampling | 64 yrs |
| Karyotype | Normal |
| Reprogramming method | Episomal vector |
| Induction method | Monolayer & chemically defined medium |
| Genetic modification | Heterozygous for the MAPT P301L mutation (CCG>CTG) |
| Genetic modification | Contains a puromycin resistance cassette (intronic) |
| Size | ≥1.5 million cells |
| Kit components | 1 vial of Neural Stem Cells (≥1.5 million cells) and 1 bottle of Neural Plating-XF Medium (30 mL) |
| Growth properties | Adherent |
| Shipping conditions | Dry ice |
| Storage conditions | Liquid nitrogen |
Frequently Asked Questions
For cortical neurogenesis, when do I switch from differentiation media (Diff-XF) to maintenance media NMM-XF?
Nnly move into maintenance media (NMM-XF) when at least 80% of your culture has differentiated to the cortical neurons.
I want to carry out imaging/confocal microscopy, how should I plate the cells?
Plate the cells on Readyset + Surebond (ax0052)
I want to passage and expand the iPSC-derived neural stem cells, what coating shall I use?
Always plate onto Surebond (ax0041) if you intend to detach the cells for continued expansion
I am not sure what coating I should use for iPSC-derived neural stem cells?
Surebond-XF - Xeno-free coating reagent needed for endpoint assays on plastic, this is the easiest method of coating, with no wash steps. Surebond + Readyset - Coating reagent needed for endpoint assays on glass Surebond- Coating reagent needed when passaging of NSCs is required
Technical Resources
References
- Dumanchin C, Camuzat A, Campion D et al. Segregation of a missense mutation in the microtubule-associated protein tau gene with familial frontotemporal dementia and parkinsonism. Human Molecular Genetics (1998)
- Hutton M, Lendon CL, Rizzu P et al. Association of missense and 5'-splice-site mutations in tau with the inherited dementia FTDP-17. Nature (1998)
- von Bergen M, Barghorn S, Li L et al. Mutations of tau protein in frontotemporal dementia promote aggregation of paired helical filaments by enhancing local beta-structure. The Journal of Biological Chemistry (2001)