Services

Knock In Mice, Genome Editing

  • Stem Cell Services

    • iPSC Generation from Patient Samples


      Our scientists have extensive experience using patient-specific PBMCs or fibroblasts to generate induced pluripotent stem cell (iPSC) lines. We have perfected both footprint-free (episomal, mRNA and other methods) and retroviral reprogramming methods for iPSC generation.

      The advantages of using our footprint-free Custom iPSC Generation Service:

      • Highly efficient reprogramming (0.2%-0.5%)

      • High quality and purity iPSCs

      • Footprint-free reprogramming of patient-derived samples: PBMCs (blood)/ fibroblast

      • Well-optimized feeder/ feeder-free protocols

      • Suitable for drug discovery and cell therapy applications

      • Comprehensive suite of iPSC characterization and differentiation to somatic lineages services (teratoma, embryoid body formation, PCR and antibody pluripotency markers detection)

    • Stem Cell Genome Editing


      ASC is one of the first “licensed” service providers of CRISPR/Cas9 and iPSC technologies, and provides the best iPSC disease modeling service on the market.

      • >98% success rate in genetically engineering hundred of iPSC lines

      • iPSC genome editing using healthy/ diseased donors

      • Customizable deliverables: Heterozygous/ homozygous mutations; Point mutations with/ without silent mutations

      • Footprint-free, feeder-free transfection and iPSC culture protocols

      • Use your iPSC lines or any of our fully characterized control iPSC lines derived from cord blood (male: ASE-9109; female: ASE-9110) or male fibroblasts (ASE-9203) that are amenable to CRISPR gene editing

      • Isogenic control lines for reliable experiments

      • Full range of downstream iPSC differentiation and characterization services

    • Safe Harbor Locus Master iPSC Genome Editing Service


      Custom “Master” iPSC Cell Line Generation using TARGATT™ technology! The TARGATT™ Master Human iPSC Line contains an attP “docking site” at the hROSA26 safe harbor genomic locus. Any gene of interest can be stably inserted at the docking site using a corresponding “attB” containing TARGATT™ donor plasmid, with guaranteed gene expression. The efficiency of this phiC31 integrase-mediated recombination is up to 100% efficiency with drug selection and up to 30% without drug selection.

      Our cell line model generation experts can also engineer your safe harbor locus knock-in iPSC lines using CRISPR/Cas9 for a multi-technology approach to generate a cell line model for your specific research needs.

    • Neural Stem Cell Differentiation


      We continually strive to improve and expand our stem cell technology platform, so researchers can leverage our expertise by using our cost-effective and reliable Stem Cell Differentiation Services for studying developmental biology, disease mechanisms, and therapeutic targets.

      • Differentiate your patient-derived iPSCs into self-renewing, multipotent NSCs and further to neurons and glial cells.
      • High differentiation efficiency and cell purity: > 95% cells express key NSC markers
      • Low passage number for robust cell culture
      • Cells fully characterized for NSC biomarkers

      We also offer upstream services to generate iPSCs from your patient-derived fibroblasts/PBMCs and downstream cell-based assays services to analyze your differentiated cells or for drug screening applications.

      Neural Stem Cell Differentiation Categories

      Astrocytes
      Differentiation
      Service

      Custom service to differentiate your iPSCs into NSCs and astrocytes progenitors, along with optimized protocols for culturing mature astrocytes.

      Astrocytes
      Differentiation
      Service

      Motor/Cortical
      Neuron
      Differentiation

      Differentiate your iPSCs to NSCs further into motor/ cortical neurons. Differentiate iPSCs from various sources into cortical neurons.

      Motor/Cortical
      Neuron
      Differentiation

      Dopaminergic Neuronal
      Cell Line

      Customized service to differentiate your iPSCs into NSCs and further into midbrain dopaminergic (DA) neuronal precursors.

      Dopaminergic Neuronal
      Cell Line

      Oligodendrocytes
      Differentiation
      Service

      Applied StemCell provides a fully customized service to differentiate your iPSCs into NScs and further into oligodendrocytes precursors.

      Oligodendrocytes
      Differentiation
      Service

    • Cardiomyocytes Differentiation


      Applied StemCell, Inc. provides a comprehensive range of service to differentiate your iPSCs into cardiomyocytes using our proprietary induction protocol and reagents. We deliver ready to use, highly pure (>90%), functional cardiomyocytes. These differentiated cells provide a highly desirable in vitro platform for high content toxicity and drug screening and as a feasible alternative to animal and embryonic stem cell models. 

      We can generate differentiated cardiomyocytes from your own pluripotent cell lines derived from healthy or disease iPSCs.

    • Hepatocytes Differentiation


      Applied StemCell provides custom differentiation of your induced pluripotent stem cells (iPSCs) into high-quality hepatocytes for developing liver disease models, discovering therapeutic treatments, drug targets, and understanding drug induced hepatotoxicity.

      Using our proprietary induction protocol and reagents, we can generate differentiated hepatocytes very efficiently and with high purity from your own healthy or disease iPSCs.

      Our Hepatocytes Differentiation Service includes:

      • Expansion of a host-derived iPSCs
      • Progenitor cell differentiation
      • Characterization of progenitor cells
      • Terminal differentiation
      • Characterization of Progenitor Cells/ Differentiated Cells by ICC (per marker)
    • Teratoma Analysis, iPSC Characterization


      Comprehensive services to fully characterize your ESC/ iPSC cell lines!

      Stringent molecular and functional assays to evaluate pluripotency for publication and to rule-out genetic aberrations are commonly required for a newly generated ES or iPSC line. We offer all services needed for a complete characterization of your pluripotent cell lines:

      • Teratoma formation analysis

      • EB (embryoid body) formation and characterization 

      • Pluripotency and lineage-specific marker immunostaining (Human/ mouse OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81, mouse Ssea-1)

      • qPCR, RNA-seq

      • Karyotyping (Chromosome counting, G-banding)

      • Germline transmission and sex determination

      ASC’s teratoma formation analysis and karyotyping service has > 97% success rate and has been acknowledged in > 30 peer-reviewed publications.

    • iPSC Culture Services


      Applied StemCell has a staff of talented scientists that can help you with the more complex to the daily laboratory tasks required of iPSC cell culture. Give your staff the freedom to focus on other project, while Applied StemCell takes care of your cell culture needs.

      We offer standard and customized cell culture services including mycoplasma testing and FBS lot evaluation.

      • iPSC expansion services
      • iPSC scale-up services
      • Neural Stem Cells, Differentiated Cells scale-up services
      • 3D-culture and morphology testing

      Please contact us to discuss your needs with our technical service specialists.

    • Custom 3D iPSC Culture Services - Organoid Modeling


      3D culture systems for use with stem cells (iPSCs) are gaining popularity as matrix grown cells have longer pluripotency, better differentiation potential, proliferation and cellular function, and are more physiologically relevant in in vitro models.

      Applied StemCell is a leader in stem cell technologies and provides custom service for 3D-iPSC culture of customer-provided iPSCs. We use our MyEZGel™ 3D-iPSC Matrix, a xeno-free nanopeptide hydrogel matrix that is specially formulated to provide an in vivo-like microenvironment to culture and generate spheroidal, physiologically relevant iPSCs.

      Applications:

      • Cell-based assay development and validation
      • Disease modeling
      • Drug discovery and screening
      • Tissue and organ engineering
      • Cell replacement therapies
    • Stem Cell Derivation


      Mouse embryonic stem cells (mESCs) are commonly used to study gene function during early embryonic development and to generate knock-in or knockout mouse models. mESCs are also a valuable tool for drug testing and high-throughput screening.

      Applied StemCell has successfully derived hundreds of mouse embryonic stem cell lines from various mouse strains including gene-targeted strains.

      We offer a service to derive novel mouse ESC lines from your own transgenic mice or from any strain-of-interest.

      We can advance your stem cell research in a timely manner.

      • ~3 weeks to derive mESC lines

      • Additional 2-4 weeks for optional characterization tests

        • Genotyping

        • Sex determination

        • Pluripotency

        • Karyotype

      • Deliverables: 2 vials of frozen, ultra-low passage (p2-p4) ESCs, each containing 1 million cells.

  • Neurotoxicity Screening

  • CRISPR/Cas9 Genome Editing

    • Cell Line Models


      Applied StemCell’s CRISPR/Cas9 cell line modeling experts have successfully engineered more than 500 distinct cell line models in > 100 distinct mammalian cell lines from different species. We use highly optimized and efficient CRISPR/Cas9 genome editing strategies and protocols to engineer or correct mutations in a variety of cell lines such as cancer cells, hard-to-transfect cells, blood lineage cells, stem cells and many more with a > 97% success rate.

      Our full service cell line editing portfolio includes:

      • Targeting vector construction and validation; cell transfection and selection; single cell cloning; expansion and cryopreservation

      • Custom deliverables: choice of homozygous and/or heterozygous clones; point mutation with/without silent mutation

      Cell Line Model Generation Categories

      CRISPR/Cas9
      Cell Line
      Service

      Our CRISPR/Cas9 service uses proprietary gRNA design and protocols to genetically modify any gene of interest in hundreds of mammalian cell lines.

      CRISPR/Cas9
      Cell Line
      Service

      CRISPR/Cas9 Cell Line
      Service - Hematopoietic Cells
      (Jurkat and TF-1)

      Applied StemCell has optimized the CRISPR/Cas9 technology to achieve the highest success rate for genome editing of human blood lineage cell lines.

      CRISPR/Cas9 Cell Line
      Service - Hematopoietic Cells
      (Jurkat and TF-1)

      CRISPR/Cas9
      Genome Editing in
      Stem Cells

      CRISPR/Cas9 genome editing in control/ patient iPSCs to engineer or correct mutations for predictive in vitro modeling of human biology and disease.

      CRISPR/Cas9 Genome
      Editing in
      Stem Cells

    • Mouse Models


      Applied StemCell is a leading CRISPR service provider, and we have genome engineered hundreds of mouse models for researchers. Our animal model portfolio includes constitutive and conditional knockout, locus-specific/ safe harbor knock-in, controlled gene expression and gene correction, to name a few.

      • Leading CRISPR service company for mouse model generation. (Nature Biotechnology (2016) 34: 893-894.)
      • Mouse models are generated in the USA in our AAALAC accredited animal facility and shipped worldwide
      • We use highly optimized protocols and ISO:9001 QMS service to generate your perfect mouse model
      • Our team of experts will discuss your project needs and suitable strategic options in detail to fit your budget and research needs; dedicated project management for milestone and final updates
      Mouse Models Categories

      Conditional Knockout Mouse
      Models

      We can generate conditional knockout mouse models to your specifications. You will have precise control over where or when your gene is knocked-out.

      Conditional Knockout Mouse
      Models

      CRISPR Mice Generation
      Service

      Custom CRISPR mouse model service using advanced design strategies, optimized protocols and validation methods to generate mouse models quickly.

      CRISPR Mice Generation
      Service

      Homologous Recombination Conditional Knockout Mouse Model and Knock-in Mouse Models

      Leverage our expertise in knock-in/ knockout mouse model generation, vector designing, ES cell targeting and mouse handling to advance your research.

      Homologous Recombination Conditional Knockout Mouse Model and Knock-in Mouse Models

      Transgenic Mice Models

      Our animal specialists can generate transgenic mouse models using bacterial artificial chromosome (BAC) or random microinjection into the pronucleus.

      Transgenic Mice Models

    • Rat Models


      With the need to physiologically relevant animals models on the rise for more predictive research findings, genetically engineered rat models are becoming increasingly popular. Get ahead of your competition by leveraging Applied StemCell’s expertise in CRISPR/Cas9 and TARGATT™ genome editing technologies and animal models engineering to generate the ideal rat model to meet your specifications and budget.

      • Global leader in genetically engineered rat models (Nature article, “CRISPR mouse model boom, rat model renaissance”)
      • Rat models are generated in the US in an AAALAC accredited animal facility using ISO:9001 quality standards
      • Detailed discussion to understand project requirements, optimal project design for precise genetic modification and dedicated project management for timely updates
      Rat Models Categories

      CRISPR Knock-In, Knock-Out,
      Conditional Knock-Out Rat

      CRISPR/Cas9 technology will be used to generate rat models that contain point mutation(s), small reporter/ gene insertions and conditional knockouts.

      CRISPR Knock-In, Knock-Out,
      Conditional Knock-Out Rat

      Bacterial Artificial Chromosome (BAC) Knock-in and Conditional Knock-In Rat Models

      We offer BAC design, cloning strategy, cloning and sequence validation of the BAC, pronuclear microinjection of BAC into rat embryos, and genotyping.

      Bacterial Artificial Chromosome (BAC) Knock-in and Conditional Knock-In Rat Models

  • TARGATT™ Genome Editing

    • TARGATT™ Site-Specific Knock-in Mouse


      Applied StemCell’s proprietary TARGATT™ knock-in mouse technology enables highly efficient and site-specific gene integration to generate transgenic mouse models. This technology uses PhiC31 integrase to insert any gene of interest into a specific docking site that has been pre-engineered into an intergenic and transcriptionally active genomic locus for guaranteed transgene expression.

      Advantages of TARGATT™ Technology for Knock-in Mouse Model Generation:

      • Safe harbor locus “large fragment knock-in” (up to 22 kb)
      • High efficiency insertion (up to 40%)
      • Single copy in an active locus: avoid gene silencing and genomic instability.
      • Germline transmitted F1 mice in 5-8 months
      • Tissue-specific/ ubiquitous, controlled/ inducible expression options

      Please contact us to discuss your project plan.

    • TARGATT™ Site-Specific Knock-in Rat


      Applied StemCell’s proprietary TARGATT™ technology enables generation of physiologically relevant transgenic rat models suitable for a variety of applications including reporter gene expression, gene knock-down, conditional gene expression and disease models. This technology uses the Phic31 integrase to mediate an irreversible integration of large transgene(s) into a preselected, safe harbor locus with guaranteed gene expression.

      • Site-specific, single copy transgene integration overcomes challenges associated with random integration
      • TARGATT™ knock-in rats in 6-9 months
      • Direct microinjection of the TARGATT™ reagents into rat zygotes

      Using the TARGATT™ technology, ASC has also developed Neural Specific Cre-Rat Lines in a Sprague Dawley rat background. Please inquire.

    • TARGATT™ Site-Specific Knock-in Cell Line Service


      Applied StemCell’s proprietary site-specific TARGATT™ technology can be used to generate stable, knock-in cell lines (large transgenes up to 22 kb) in many cell lines, including stem cells, very efficiently and quickly.  Knock-in is mediated by PhiC31 integrase at a pre-engineered "docking site" in an intergenic, transcriptionally active genomic locus (safe harbor locus) for guaranteed gene expression without disruption of internal genes. This technology allows only a single-copy integration with very high efficiency without or without clonal selection.

      Use our TARGATT™ technology to generate your "Master" cell lines, reporter/tag lines, for iPSC generation, conditional gene expression models and more.

      Also, try our ready-to-use TARGATT™ Master Cell Lines to knock-in transgenes in your own lab.

    • EvolvOne™ TARGATT™ Protein Screening Platform


      The TARGATT™ technology enables faster and efficient site-specific integration of large DNA fragments in cell lines. This technology offers an ideal platform for generating stable isogenic cell line libraries for mammalian cell display-mediated antibody engineering, protein evolution screening, mammalian two-hybrid (M2H) screens and more. It allows only a 1:1 variant-to-cell ratio, and guarantees uniform and consistent expression of the gene/protein for efficient screening.

      • Single copy knock-in: 1 cell, 1 docking site, 1 inserted transgene
      • Site-specific knock-in: avoids problems associated with random insertion
      • Isogenic cell library: comparison of phenotype against identical baseline
      • Cost-effective and efficient library construction: no virus packaging time and resources
      • BSL1 compatible
  • Bioproduction

    • CHO Cells


      TARGATT™ Chinese Hamster Ovary (CHO) Cells for Recombinant Proteins and Antibody Bioproduction

      Applied StemCell has created Master TARGATT™ CHO cell lines for the rapid creation of new lines for high level protein and antibody expression. The TARGATT™ CHO master cell lines yield >2.5g/L of recombinant proteins in a 2-week fed-batch shake flask expression system and is easily scalable for large scale bioproduction. 

      Our Master TARGATT™ CHO cell lines exceed the capabilities of the traditionally made CHO antibody production cells and also reduces the cost of bioproduction and manufacturing. Our technology makes CHO antibody production feasible and economical for companies and projects of all sizes.

      Please let us know how we can help you with your CHO bioproduction project!

    • Transgenic Animals - Rabbit


      TARGATT™ Rabbit for Biopharming and High-Yield Recombinant Protein and Antibody Production

      Animal bioproduction uses transgenic animal mammary gland as a bioreactor for the production of recombinant proteins. Animal bioproduction, compared to CHO bioproduction, has the advantages of lower upfront and maintenance costs; is easy to contain, control and transport; involves faster development processes and no scale-up issues; and has a unique low-cost bulk holding stage (frozen milk). However existing technological methods such as random integration to create transgenic animals have many limitations. Applied StemCell has overcome these limitations with our proprietary TARGATT™ technology to reproducibly, and consistently, control the knock-in locus, copy number and expression of the target protein in transgenic animals.

  • Lentivirus Stable Cell Line Generation

  • DNA Cloning & Virus Packaging

    • Virus Packaging (Lentivirus and Retrovirus)


      Applied StemCell provides lentiviral and retroviral custom virus packaging services so you can focus your efforts on other aspects of your project. We also provide custom virus packaging for efficient delivery of CRISPR/Cas9 components for in vivo and in vitro models.

      • Rapid, custom lentivirus or retrovirus virus packaging

      • Ready-to-transduce viral particles provided to you

      • High quality, high titer viral particles in 10 days

      • VSV-G pseudotyped viruses that exhibit broad tropism across a range of cell types

      • High titer amplification of viruses, up to 10^9 infectious units per ml

      • Accurate viral titers quantified by qRT-PCR

      • Confirmation by DNA sequencing

      • Large scale production, clonal expansion and cryopreservation

    • CRISPR / Cas9 Vector Design & Construction


      Applied StemCell is one of the earliest licensees and provider of CRISPR/Cas9 genome editing services. We are experts in designing and cloning components for CRISPR-based gene editing in cell lines and animal models using benchmark strategies and optimized protocols. Our scientists have designed > 1000 CRISPR projects for researchers worldwide, and we use the latest sequencing and next generation sequencing (NGS) protocols to validate your gRNAs and to ensure high efficiency gene modification in your gene of interest.

      We offer CRISPR/Cas9 vector design, construction and gRNA validation for:

      • Cell line gene modification
      • Microinjection into embryos for animal model generation
      • Virus packaging
    • Vector Design / Cloning Service


      Applied StemCell has extensive expertise in anything DNA-related. We can develop the best strategy to tackle your cloning projects, and find solutions to all your technical problems.

      • Comprehensive services for genome engineering for in vivo and in vitro applications
      • Restriction fragment cloning
      • Design and validation of CRISPR/Cas9 components: gRNA design and in vitro validation by next generation sequencing (NGS)
      • Gene targeting vectors for homologous recombination
      • Design and construction of vectors for transient mammalian transgene expression
      • Bacterial artificial chromosome recombineering for large fragment insertion
      • Design of RNAi and inducible vectors, gene tagging, and site-directed mutagenesis
      • Vector design for generation of random transgenic cell line and animal models
    • Bacterial Artificial Chromosome (BAC) Design and Cloning


      Bacterial Artificial Chromosomes (BACs) are ultra-low copy vectors that can hold up to 300 kb of genomic fragments, making them ideal vectors for introduction of entire genes including the regulatory regions for disease modeling with transgenic animals. BACs are traditionally difficult to modify with restriction enzymes and ligases because of their large size. Applied StemCell uses homologous recombination to introduce precise changes into BACs for your specific experimental needs. Virtually any desired modification can be introduced into a BAC, including insertion of reporter genes, point mutations, Lox-STOP-Lox conditional modifications, and more.

      Our custom BAC services include:

      • Design
      • Cloning
      • Validation

      Applied StemCell can also help you create transgenic rats using BACs.

  • CRO Services

    • Drug Toxicity and Efficacy Testing


      Make Informed Go-No-Go Decisions Early in Your Drug Development Process!

      Leverage Applied StemCell’s unique expertise in CRISPR/Cas9 and induced pluripotent stem cell (iPSC) technologies to engineer predictive and reliable in vitro disease models as an alternative to animal models for preliminary drug screening.

      Our ISO:9001 certified service platform offers full flexibility in choosing assay modules with a wide-range of functional endpoints for early-stage in vitro screening of preclinical drug candidates. We have a comprehensive cell-based test battery from which you can choose assays for efficacy, safety or target discovery that suit your therapeutic pipeline. We provide the scientific expertise, accurate and efficient screening to help you make informed decisions about your small/ large molecules early in your drug development process.

    • Custom In Vivo Assay Services


      As a long-standing leader in genetically engineered animal models, Applied StemCell also offers a comprehensive custom research solutions platform for downstream assays in mouse and rat models. Our state-of-the-art vivarium is equipped with automated cages & devices for behavioral assessments, sample collection and in vivo measurements (ECG, EEG) of the animals. We have a multidisciplinary team of expert scientists who can design a comprehensive project plan to fit any requirement/stage of your research pipeline and for drug discovery and screening.

      • Designing and engineering research animal disease models
      • Adoptive transfers/transplantation
      • In vivo functional screening assays
      • End-of-study in vitro assays
    • Cell Line Irradiation Service


      Applied StemCell offers gamma-irradiation service for irradiating cell lines (patient-derived, pooled, genome edited/ corrected cell lines) using a Nuclear Regulatory Commission (NRC)-approved Gamma-emission source to irradiate cells at a constant dose rate and to any desired extent of radiation. Irradiated cell line models are novel toolkits for applications such as generation of co-culture models using non-dividing/ feeder cells; to study mechanisms underlying cancer progression; wound healing and inflammatory responses after irradiation; screen drugs for enhancing irradiation sensitivity of tumor cells; and for xenograft tumor models.

      Our Cell Line Irradiation Service includes:

      1. Cell line expansion/ harvesting
      2. Cell line Irradiation (customized extent of irradiation)
      3. Cryopreservation
    • Custom Services


      To speed up your research and development; we provide pre-clinical development services (CRO) and support. Our custom services will allow you to focus on the main aspects of your research while we develop the interim paradigms for you.

      Custom Services Categories

      Fibroblasts Immortalization Service

      Simian virus 40 T antigen to immortalize human fibroblasts is the foundation of Applied StemCell’s Fibroblasts Immortalization Service platform.

      Fibroblasts Immortalization Service

      Custom Primary Cell Isolation Service

      Employing optimal isolation and culturing techniques, Applied StemCell will isolate primary cells from organs or tissues for your ideal cell model.

      Custom Primary Cell Isolation Service

      FFPE Service

      Preserve your cells by FFPE (Formalin-Fixed Paraffin-Embedded) to create a reference standard, use for IHC, ISH or other applications.

      FFPE Service

Have Questions?

An Applied StemCell technical expert is happy to help, contact us today!