Custom cell line gene modification; KI; Timeline: 3-5 months
Référence ASC-7102
Conditionnement : 1service
Marque : Applied StemCell
Automated High-throughput Protocols for the Genetic Modification of Your iPSC Line of Choice
We can engineer your control and disease iPSC lines or choose from one of our well-characterized master iPSC lines derived from fibroblasts (male: ASE-9211; female: ASE-9209), cord blood, or PBMCs, which have proven CRISPR gene editing and differentiation potential.
Faster Timelines with Automated High-throughput Protocols | |||
Project type | Conventional Protocols | ASC’s Optimized High-throughput Protocols | Improvement in Delivery Times |
Knockout (KO) | 12-20 weeks | 6-8 weeks | 60% |
Point Mutation (Single Nucleotide Polymorphism or Variant) | 12-20 weeks | 6-8 weeks | 60% |
Knock-in (Reporters/ Tags, Large Transgenes) | 12-24 weeks | 10-15 weeks | 20-30% |
A Variety of Other Modifications: Standard & Complex
Correct/engineer mutations or introduce a variety of genetic modifications in iPSCs:
- Gene knockout: gene disruption or site-specific large fragment knockout (>10kb)
- Gene insertion: reporter gene/ tag insertion, small fragment insertions, SNV/ point mutations
- Inducible gene expression/ gene overexpression models
- Gene fusion (translocation, inversion, etc)
Don't limit yourself to only the standard modifications. Ask us about: Multiplexed genome editing; conditional knock-in, gene fusion, and other models that you would like for your projects.
And…. We offer Customized Deliverables
- Choice of heterozygous or homozygous mutations
- Footprint-free genome editing – Ex. Single nucleotide variant (SNV; point mutation) engineering without silent mutations for regulatory compliance
- Specific genetic or safe harbor locus
CRISPR Knock-In Projects
Project 1:
Goal: Knock-in of 1 bp at the AAVS1 locus using the ASE-9211 Master iSPC Line by CRISPR/Cas9 technology
Knock-In Strategy for AAVS1 (1bp insertion)
Figure 1: Knock-in strategy for 1bp insertion in the AAVS1 locus of the ASE-9211 Master Cell Line.
Genotyping Clone #6
Figure 2: Sequencing chromatogram of iPSC line with 1bp insertion in the AAVS1 locus (top: Clone #6) compared to the Parent line, ASE-9211 (bottom).
Project 2:
Goal: Knock-in of 150bp at the AAVS1 locus using the ASE-9211 Master iPSC Line by CRISPR/Cas9 technology
Knock-In Strategy for AAVS1 (150bp insertion)
Figure 3: Knock-in strategy for 150bp insertion at the AAVS1 locus of the Master iPSC Line.
Genotyping Positive Clone #21
Figure 4: Sequencing chromatogram showing the ~150bp insertion at AAVS1 locus.
CRISPR Knockout Projects
Project 3:
Goal: 1bp deletion in the AAVS1 locus using the ASE-9211 Master Cell Line by CRISPR/Cas9 technology
Figure 5. Sequence chromatogram of iPSC line with 1 bp deletion (AAVS1-1bp DEL; bottom) compared to wild type (WT; top).
Figure 6. Sequence alignment between the 1 bp deletion iPSC line (AAVS1-1bp DEL; bottom) and wild type (WT; top).
Project 4:
Goal: 1000bp Deletion in the AAVS1 locus using the ASE-9211 Master Cell Line by CRISPR/Cas9 technology
Figure 7. AAVS1 wild type (WT) sequence showing gRNA cut sites and position of 1007 bp (~1000 bp) deletion (sequence in red).
Figure 8. AAVS1 WT chromatogram showing sites of ~1000 bp deletion (sequence in red). Top: Sequence for 5’ deletion site; Bottom: Sequence for 3’ deletion site.
Figure 9. Sequence chromatogram of iPSC line with ~1000 bp deletion in the AAVS1 locus.
Only a few NIST projects are listed, if you would like to learn more, contact us today.
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