Genome Editing Services for Cell Lines

CRISPR Service, Cell Line Services

CRISPR Genome Editing for Cell Lines

Genomic Engineering in cell lines is a versatile tool for studying gene function, designing diseases models, biopharmaceutical research, drug discovery and many other applications. Applied StemCell is excited to present our cell line services including genome engineering, gene editing, and knock-in cell lines. To date, methods of gene insertion in human stem cells (viral vectors, transposons, or plasmids) are mostly random insertions subject to silencing and disrupting internal genes, inefficient, multiple copies, loss of expression after differentiation, and have limitations with the insert DNA size. Nuclease-based methods, particularly the CRISPR-Cas9 method, offer an efficient way of controlled gene insertion. However, creating double strand breaks also can have negative consequences including cellular toxicity, off-target recombination, and sequence alterations near the target site. Visit our CRISPR services page for more specific information on CRISPR-Cas9 Gene Editing services.

TARGATT Genome Editing for Cell Lines

Site-specific integrases, such as the TARGATT™ method, offer an attractive method for precise gene addition in human stem cells. Integrases such as phiC31 and Bxb1 catalyze recombination between two non-identical sites, attB and attP. This feature, along with lack of a corresponding excisionase activity, makes the recombination reaction unidirectional and efficient. With this method, “master” human stem cell lines were constructed to contain a “docking site” of attPs. The attP sites were directed to a “safe harbor” genomic locus that was shown to provide robust transcription in all cell types previously in the mouse study. These docking-site-ready (DSR) cells can be used to insert any genes of interest in a fast, simple and site-specific manner with high efficiency (100% with selection). This new DSR master cell line system combines integrases with novel open chromatin loci and ensures efficient, uniform, and robust gene expression.

Comprehensive Technology Platform for Gene Editing

Methods Technical Advantage
TARGATT™ phiC31 integrase Site specific integration in "Safe harbor locus"(ROSA26)
High efficiency(up to 40%)
Works for large fragment knock-in(-22kb)
Insert promoter of choice for gene: overexpression and inducible expression
Works independent of cell division
CRISPR / Cas9 High specificity
High frequency for Knockout, point mutation
Large DNA knock-in up to 5kb
Generate homozygous or heterozygous modified cell lines
Possible Cell Line Engineering Projects (selected):
  • Knock-in/knock-out cell lines
  • Gene replacement
  • Gene editing including single base mutations
  • Gene tagging
  • Gene Therapy
  • Promoter modifications
  • Mutagenesis
  • Removal of viral sequences
  • Stable cell lines / immortalizatioin 
… and many more. Please contact us to discuss your project.
 
Applications (selected)
  • Recombinant protein production in CHO cells
  • Disease model in human cell lines for drug discovery
  • Gene therapy in diseased cell line
  • Generation of a specific cell line with TARGATT™ docking site as a master cell line for site-specific gene knock-in reporter cell line for gene function studies