CRISPR/Cas9 Genome Editing

CRISPR Genome Editing

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.

 schematic-crispr-cas9-technology

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 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.

CRISPR applications include: Functional genomics, disease modeling, target identification and validation for drug discovery and screening, and many more.

  • Cell Line Model Generation

    CRISPR Genome Editing for Cell Lines

    We offer full-service for gene editing in any mammalian cell and targeting any gene of interest. Our CRISPR cell line editing portfolio includes > 300 cell line models generated from > 100 distinct parental cell lines, including hard-to-transfect cell lines, blood lineage cell lines, cancer cell lines and stem cells.

  • Mouse Models

    Applied StemCell's comprehensive CRISPR mouse model generation service includes a broad range of gene modification portfolios suitable for all preclinical research applications, as well as phenotype analysis. Applied StemCell, Inc. has an AAALAC accredited animal facility and all mice and rats are generated within the USA.

  • Rat Models

    Applied StemCell offers full range of CRISPR/Cas9 engineered transgenic rat model generation service with a variety of genetic modifications suitable for all preclinical research applications, including research applications in the neuroscience and cancer space. All animal models are engineered in our US facility under strict regulatory protocols.

Choose the right gene editing technology:

Applied StemCell also uses a propriety site-specific integrase-based gene editing technology, TARGATT™ for large fragment (up to 20 kb) knock-in in safe harbor locus. The complementary TARGATT™ and CRISPR/Cas9 technologies can be used together to generate advanced cell line and animal models very efficiently and effectively.

Project Purpose

CRISPR/Cas9

TARGATT™

Knock-Out (KO)

Yes

 

Point Mutation

Yes

 

Conditional KO

Yes

 

Knock-In

(<200 Nucleotide ssODN Donor)

Yes

 

Knock-In Transgenes in

Safe Harbor Loci (>2kb)

Challenging

(but limitations on size)

Yes

 (up to 20kb)

Knock-In

 (Plasmid DNA)

Challenging

(but limitations on size)

Yes

 (2 steps: KI docking site; KI transgene)