iPSC services: Induced Pluripotent stem cells (iPSCs) are a type of pluripotent stem cells that can be reprogrammed from adult somatic cells, derived from healthy or diseased patients. These pluripotent stem cells provide a limitless supply of patient-derived samples to develop in vitro preclinical models of diseases, that represent the genetic and epigenetic features of a disease better. As well, iPSCs are amenable to genome editing which has been a stumbling block for developing research models using primary cells lines.
Applied StemCell (ASC) provides a wide range of integrated embryonic stem cell (ESC) and iPSC services, including iPSC generation/ ESC derivation, disease modeling/ correction using CRISPR/Cas9 and our proprietary TARGATT™ genome editing technologies, a complete panel of stem cell characterization (teratoma formation assays, pluripotency marker assays and more), and stem cell differentiation.
ASC is one of the earliest licensees of both the iPSC reprogramming factors from iPS Academia, Japan and the CRISPR/Cas9 technology from the Broad Institute. Using optimized, feeder-free protocols, we are experts in handling and maintaining healthy, pluripotent iPSCs at every stage of your project(s). ASC’s acclaimed CRISPR/Cas9 modified iPSC services for in vitro genetic disease modeling has even been featured in the article, “CRISPR/Cas9-transforming gene editing in drug discovery labs” (Comley, et al., 2016) published by the peer-reviewed magazine, Drug Discovery Weekly.
Applied StemCell offers comprehensive preclinical neurotoxicity and neuroprotective compound testing for potential drug candidates using human iPSC-differentiated neural lineage cells
The CRISPR Genome Editing technology is a versatile tool for efficient gene modification of nearly all types of cells and has gained popularity in just a few years due to the simplicity of design and delivery of its RNA-probes, high specificity and efficiency, ease of implementation, cost and turnaround time. The CRISPR/Cas9 technology utilizes Cas9 endonuclease to introduce sequence-specific double stranded DNA breaks (DSB) using appropriate guide RNAs (gRNAs) and DNA repair mechanisms, the non-homologous end joining repair (NHEJ) or the homology directed repair (HDR), to precisely edit genes. This technology although revolutionary, is still in its infancy and its outcome can be tainted by off-target mutations resulting from NHEJ-mediated indels in non-targeted sites after indiscriminate Cas9 cleavage and unspecified epigenetic factors at the targeted site.
Applied StemCell is one of the earliest licensees of this technology (from the Broad Institute), and we have gained extensive experience in using the CRISPR technology by having engineered hundreds of animal and cell line models for biomedical research. We offer ISO:9001 quality management system certified custom service to introduce a variety of genetic modifications in cell lines (including stem cells), mouse and rat models. Visit our CRISPR services page for more specific information on Applied StemCell's acclaimed CRISPR-Cas9 Gene Editing services, and to leverage our expertise in advancing your research and discoveries.
Genetic modifications available through our CRISPR gene editing platform:
Gene knockout, Point mutation knock-in, gene insertion in any locus, including safe harbor locus (large fragment insertion, reporter gene knock-in, gene replacement), conditional knockout/ knock-in models, conditional/ inducible gene expression models.
Functional genomics, disease modeling, target identification and validation for drug discovery and screening, and many more.
Recent advances in gene editing technologies have greatly increased our ability to make precise changes in the genome of cells or embryos and have eliminated major stumbling blocks in the generation of more sophisticated transgenic animal and cell line models.
Applied StemCell’s proprietary, TARGATT™ that allows for the site-specific integration of large DNA fragments, more efficiently and faster. The novel TARGATT™ technology uses the PhiC31 bacteriophage integrase to mediate an irreversible recombination between a pre-engineered “attP” docking-site and an attP recognition-sequence on the donor vector “attB”, for stable gene integration. The attP docking sites can be engineered into a specific or intergenic, transcriptionally active genomic locus (safe harbor locus), using CRISPR/Cas9. The transgene integration is site-specific, stable and with guaranteed expression.
TARGATT™ gene editing platform is very versatile and can be adapted to generate large fragment knock-in animal or cell line models from various species. This technology circumvents problems associated with random integration such as position effect, and gene silencing or instability due to integration of multiple copies of the transgene.
With Applied StemCell’s two complementary gene editing technologies, TARGATT™ and CRISPR/Cas9, we can generate a variety of advanced genetic modification in animals and cell lines to expand the scope of biomedical research and discoveries.
Advantages of TARGATT™ Technology:
- High integration efficiency (up to 40%)
- Large transgene knock-in (up to 22 kb)
- Reduced time and cost
- Guaranteed, high level expression of the transgene
- Site-specificity allows a precise comparison of the effects of the transgenes among different lines
- Site-specific knock-in at pre-selected locus overcomes challenges associates with random integration:
- Eliminates position effect
- Integration at intergenic region ensures that no internal genes are interrupted
- Single copy gene integration eliminates repeat-induced gene silencing and genomic instability
- Transgene overexpression models (Ex. For bioproduction)
- Humanized animal models (large gene insertion)
- Reporter gene insertion models
- Inducible expression models (Example: Tet-regulatory systems)
- Cre - driver animal models - We can generate Cre animal models using promoters listed below or ANY promoter provided by customers (with available sequence) using our versatile TARGATT™ technology:
Lentivirus stable cell line generation in hard-to-transfect cell lines, non-dividing and primary cells: Gene knockout and gene overexpression cell line models
Customized DNA cloning service for vector design and cloning strategies.
With decades of experience our Molecular Biology Team has developed an exceptional expertise in anything DNA-related. We can help you develop the best strategy to tackle your cloning projects and find solutions to all your technical problems.
Cloning of amplified DNA fragments RNAi and inducible vectors Gene targeting vectors for homologous recombination Site-specific mutagenesis Bacterial Artificial Chromosome (BAC) Recombineering Tagging of genes De novo gene synthesis
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.
Media development Animal phonotype, behavior analysis (Mouse / Rat) Mycoplasma testing Bio assay development FBS testing Loci analysis Animal injections
The traditional CHO antibody production method and animal bioproduction methods are very inefficient because it involves random gene insertion and forced amplification of transgenes. Many factors adversely affect bioproduction levels. These factors include insertion sites effects (position effects), DNA repeat-induced gene silencing and genomic instability, altered regulation or interruption of endogenous genes.
Applied StemCell Bioproduction platform uses its proprietary TARGATT™ technology to express recombinant proteins that circumvents the disadvantages associated with traditional random insertion techniques. It features two efficient and high yielding methods: TARGATT™ CHO Master Cell Lines and TARGATT™ Rabbit for low-cost, high-yield and consistent protein expression that is easily scalable for large scale bioproduction. With our comprehensive bioproduction services and our extensive experience in engineering hundreds of cell lines and animal models, we can help you traverse your drug development process without stress. The TARGATT™ bioproduction platform is suitable for companies of all sizes and drug development applications.
Applied StemCell offers multiple products for embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) generation, stem cell characterization and stem cell differentiation. See below for our reagents and kits for the generation of iPS cell lines, characterization of pluripotent stem cells, and differentiation of pluripotent stem cells. We also provide individual components for our stem cell generation, stem cell characterization and stem cell differentiation kits.
- CRISPR/Cas9 Genome Editing
TARGATT Genome Editing (Knock-in)
Isogenic Cell Lines for Drug Dicovery: Isogenic cell lines represent a unique toolset for studying the impact of genotype on cellular phenotype.
Isogenic Cell Lines for Drug Dicovery: Isogenic cell lines represent a unique toolset for studying the impact of genotype on cellular phenotype. An isogenic cell line is a cell line that has been engineered from a parental line through the introduction of a targeted gene mutation. In doing so, the parental cell line inherently becomes a control line to which the engineered line can be referenced. Isogenic cell line pairs can be incredibly useful as tools for studying cellular biology, such as the impact of genotype on cellular phenotype, as well as for parallel, high throughput screening to enable the discovery of therapeutic compounds that exhibit genotype-specific toxicity. Applied StemCell offers a number of homozygous isogenic cell lines.
For genome engineering applications, we also offer cell lines that have been modified to enable more efficient genome editing, such as our Cas9-expressing Jurkat line, and hiPSC TARGATT™ Master line.
In addition to our off-the-shelf products, we offers custom cell line engineering services for the generation of knock-out, point mutation, knock-in, inducible, and reporter cell lines.
Applied StemCell Inc. provides a variety of disease human/mouse primary cells and normal human primary cells. Our human primary cells, cDNA, RNA collections are obtained from patients who have been clinically diagnosed with specific disorders.
Applied StemCell's disease cell lines include: neurological disorders, oncogenic disorders, muscular disorders, genetic disorders, autoimmune disorders, endocrine disorders, congenital disorders, cardiovascular disorders, metabolic disorders, degenerative disorders, blood disorders.
- Normal and diseased fibroblasts, primary cells
- isolated RNA
- Immortalized Fibroblasts