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Genome Engineering and iPSC Center

St. Louis, Missouri, US

The Genome Engineering and IPSC Center (GEiC) was formed by the consolidation of two pre-existing cores, the Genome Engineering Center and the Induced Pluripotent Stem cell (iPSC) core, both established by the Department of Genetics in the past few years. These two Centers were established to facilitate functional genomic studies through the use of patient-derived iPSCs and the generation of modified cells and organisms using genome editing technologies. The recent merging of these Centers reflects the new opportunities afforded by uniting these powerful new technologies. The GEiC includes a consultative service whereby investigators are able to query experts in genomic engineering technologies and in generation and differentiation of patient-derived induced pluripotent stem cells (iPSCs).

Papers Citing Our Services

  • Discovery of a proteinaceous cellular receptor for a norovirus. Orchard et al., Science... Show more »

The Genome Engineering and IPSC Center (GEiC) was formed by the consolidation of two pre-existing cores, the Genome Engineering Center and the Induced Pluripotent Stem cell (iPSC) core, both established by the Department of Genetics in the past few years. These two Centers were established to facilitate functional genomic studies through the use of patient-derived iPSCs and the generation of modified cells and organisms using genome editing technologies. The recent merging of these Centers reflects the new opportunities afforded by uniting these powerful new technologies. The GEiC includes a consultative service whereby investigators are able to query experts in genomic engineering technologies and in generation and differentiation of patient-derived induced pluripotent stem cells (iPSCs).

Papers Citing Our Services

  • Discovery of a proteinaceous cellular receptor for a norovirus. Orchard et al., Science 2016
  • Blood Group O–Dependent Cellular Responses to Cholera Toxin: Parallel Clinical and Epidemiological Links to Severe Cholera. Kuhlmann et al., Am J Trop Med Hyg 2016
  • Clec16a is Critical for Autolysosome Function and Purkinje Cell Survival. Redmann et al., Sci Rep 2016
  • Multiple Domains of GlcNAc-1-Phosphotransferase Mediate Recognition of Lysosomal Enzymes. Van Meel et al., JBC 2016
  • A Puromycin Selectable Cell Line for the Enrichment of Mouse Embryonic Stem Cell-Derived V3 Interneurons. Xu et al., Stem Cell Research & Therapy 2015
  • A Noncanonical Autophagy Pathway Restricts Toxoplasma gondii Growth in a Strain-Specific Manner in IFN-γ-Activated Human Cells. Selleck et al., MBio 2015
  • A calcium-dependent protease as a potential therapeutic target for Wolfram syndrome. Lu et al., PNAS 2014

Genome Engineering and Stem Cell Technology Papers by Our Scientists

  • Donor plasmid design for codon and single base genome editing using zinc finger nucleases. Pruett-Miller, et al., Methods Mol Biol 2015
  • Gene editing using ssODNs with engineered endonucleases. Chen et al., Methods Mol Biol 2015
  • Preface. Chromosomal mutagenesis. Pruett-Miller, Methods Mol Biol 2015
  • High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs. Duda et al., Nucleic Acids Res 2014
  • Nuclease-mediated gene editing by homologous recombination of the human globin locus. Voit et al., Nucleic Acids Res 2014
  • Genome editing in mouse spermatogonial stem/progenitor cells using engineered nucleases. Fanslow et al., PLoS One 2014
  • Targeted correction of RUNX1 mutation in FPD patient-specific induced pluripotent stem cells rescues megakaryopoietic defects. Connelly et al., Blood 2014
  • A calcium-dependent protease as a potential therapeutic target for Wolfram syndrome. Lu et al., PNAS 2014
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Cell Lines
Price on request

Custom iPSC Line Creation

CRISPR nucleases can be used to create iPSC lines with specific genomic modifications (as above). The capability to genetically incorporate (or correct) disease-causing point mutations in patient-derived iPSC lines will be invaluable for elucidating disease mechanisms through functional genomics, for... Show more »

Custom iPSC Line Creation

CRISPR nucleases can be used to create iPSC lines with specific genomic modifications (as above). The capability to genetically incorporate (or correct) disease-causing point mutations in patient-derived iPSC lines will be invaluable for elucidating disease mechanisms through functional genomics, for identifying therapeutic agents and for the development of new cell-based therapies. The GEC has now been merged with the Induced Pluripotent Stem cell (iPSC) facility, which banks patient fibroblasts, generates iPSCs, and develops iPSC differentiation schemes. This allows for a seamless transition from patient derived iPSC generation to the engineering of modified iPSC lines. The GEP will have access to all the capabilities of this newly combined Center.

Donor design and creation services

To produce user-defined knockin mutations (e.g. incorporation of disease-causing point mutations), a donor substrate (either DNA fragment or oligonucleotide) must be co-delivered with the CRISPR nuclease and incorporated into the locus by homology directed repair. We offer donor plasmid and donor oligonucleotide design and validation services in addition to the requisite CRISPR nucleases.

  • For each knockin project, a GEiC scientist will discuss your overall project goals and design appropriate plasmid or oligonucleotide donors.
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mRNA Synthesis
Price on request

CRISPR nuclease mRNA production that is ready for embryo injection

CRISPR nucleases have successfully been used to create novel mouse and rat transgenic models. Direct injection of mRNA encoding Cas9 and the gRNA into one-cell stage embryos results in rapid and efficient generation of model animals with a wide variety of... Show more »

CRISPR nuclease mRNA production that is ready for embryo injection

CRISPR nucleases have successfully been used to create novel mouse and rat transgenic models. Direct injection of mRNA encoding Cas9 and the gRNA into one-cell stage embryos results in rapid and efficient generation of model animals with a wide variety of modifications including gene deletions, point mutations, conditional knockouts, epitope and fluorescent reporter gene tagging and multiplex modifications. Although, AMRF investigators are primarily utilizing rodent models, most other organisms can be modified using CRISPR/Cas9 methods because the technology is species-agnostic.

  • CRISPR nuclease mRNA production
    • We synthesize both the Cas9 mRNA and gRNA in a form that is ready for direct embryo injection.
  • Deliverables – 2 week turnaround time
    • gRNA at >300ng/ul concentration
    • Cas9 mRNA at >100ng/ul (14ug total)
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CRISPR/Cas9 vector generation
Price on request

Multiple backbones are available (e.g. separate Cas9 and gRNA backbone, all-in-one Cas9/gRNA backbone, all-in-one Cas9-GFP backbone, etc)

  • CRISPR nuclease design*
    • We do a full bioinformatics workup for each target that includes an off-target analysis, SNP check, and recommendations for gRNA placement within your region of... Show more »

Multiple backbones are available (e.g. separate Cas9 and gRNA backbone, all-in-one Cas9/gRNA backbone, all-in-one Cas9-GFP backbone, etc)

  • CRISPR nuclease design*
    • We do a full bioinformatics workup for each target that includes an off-target analysis, SNP check, and recommendations for gRNA placement within your region of interest.
  • CRISPR nuclease assembly and validation
    • We prepare two gRNA plasmids and functionally test them for each project. We introduce the CRISPR nuclease into cells and use our validation assay to demonstrate that the CRISPR nuclease is cutting the desired endogenous, chromosomal target site. We only charge for a project after we have successfully identified at least one active CRISPR nuclease.
  • Deliverables – 3-4 week turnaround time
    • Plasmids containing the gRNAs and the Cas9 expression plasmid
    • The primers used in validation assay to detect CRISPR nuclease activity.
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Genotyping Services
Price on request

Genotyping services include assay design, targeted deep-sequencing, STR profiling, and zygosity analysis.
- Animal genotyping (e.g. edited F0 screening)
- Transgene integration
- STR analysis
- Edited cell pools

Genotyping services include assay design, targeted deep-sequencing, STR profiling, and zygosity analysis.
- Animal genotyping (e.g. edited F0 screening)
- Transgene integration
- STR analysis
- Edited cell pools

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Biology
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Plasmids and Vectors
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Biochemistry & Molecular Biology
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DNA
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DNA Services

DNA Services

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Nucleic Acid Services
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Cells and Tissues
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RNA
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2018-05-25 17:09:33 +0200

Net Promoter Score of 10 received for DNA.

Additional Ratings: satisfaction with deliverable: 10, satisfaction with timeliness: 10.

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