NIST Genomic DNA Reference Standards & Engineered Human iPSC Line
Applied StemCell (ASC) became a member of the National Institute of Standards and Technology (NIST) Genome Editing Consortium in 2018. Its “Master” iPSC Line is being utilized to develop measurements and standards for genetically engineered cells that can be potentially be used by gene and cell therapy, NGS, biotechnology, and diagnostic companies as well as other key industry players as benchmark data.
ASC is a leading induced pluripotent stem cell (iPSC) service provider that has over 13 years of expertise in iPSC generation, characterization, precise genome editing, differentiation, and analysis in both cells and animals. With our well-established iPSC platform, we have shared our expertise and completed a series of CRISPR-iPSC genome editing projects for the NIST program. ASC designed and executed various insertion and deletion projects using our well-characterized Master iPSC Line, ASE-9211. Our scientists have confirmed the desired mutations and delivered the genetically modified iPSCs, and we expect to see additional characterization data from the other members in the near future.
The genomic DNA reference standards and the engineered human iPSCs are now available for purchase. Explore our product catalog below or contact us today to learn more.
Products and Services
Viewing 1-16 of 16 products
ASC provided its proprietary iPS cell line, ASE-9211 as the starting material for all projects. Single cell cloning, screening, and genotyping by PCR and sequencing was completed by ASC.
CRISPR Knock-In Projects
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).
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
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).
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.