Cell Line Model Generation

CRISPR Genome Editing

CRISPR Genome Editing for Cell Lines

CRISPR/Cas9 edited cell line models are great in vitro tools for studying gene function, designing disease models, for drug discovery and high throughput compound screening. At Applied StemCell, we have experience in editing > 100 distinct mammalian cell lines from different species, and have engineered more than 300 cell line models. We can genetically modify hard-to-transfect cells, blood lineage cells, slow growing cell lines, adherent/ suspension cells, stem cells and correct mutations in disease cell lines with a >97% success rate.

Cell Type Modification Type

Selected Cell Lines From > 100 distinct parental cell lines engineered

A-549     BEAS-2B    BT-474     HaCaT     HBE Huh7     MCF-10A     OCCM-30     RPE-1     SK-MEL-31     Tert-RPE     U-2 OS     786-O     CHLA-10     A-375     Gist-T1     DLD-1     HCT-116     HEK293     HEK293T     HeLa     HepG2     4T1     C2C12     cTEC     ES Cells     iPSCs     MWCL-1     BCWM-1     H929     Jurkat     K562     KHYG-1     LAD2     MM.1s     MWCL-1     NCI-H929     T2 cells     TF-1     HT1080     HT29     KBM-7     KN12-Luc     LnCap     MDA-MB231     NCI-H2228     RKO     TC32     SCC35     SH-SY5Y

 

 

 

 

Cancer Cell Lines

KO

Point Mutation

Insertion

Human iPSCs

KO

Point Mutation

Insertion

Human ES Insertion
Primate iPSCs Insertion
Human Primary Cells

KO

Insertion

Human Fibroblasts Point Mutation
Mouse Fibroblasts Point Mutation
Rat Thyrocytes KO

Full service cell line editing porfolio includes:

  • Choice of zygosity: homozygous and/or heterozygous clones
  • Choice of silent mutation at targeted site for point mutation insertion
  • Dedicated project management for detailed milestone and final reports


Some examples of cell line engineering projects include but are not limited to:

  • Knock-in/knockout modifications
  • Gene replacement
  • Gene overexpression
  • Conditional / inducible gene expression
  • Gene editing/ correction, including single base mutations
  • Gene tagging
  • Gene therapy
  • Promoter modifications
  • Gene fusion/ translocation
  • Removal of viral sequences
  • Stable cell lines / immortalization 

Please contact us to discuss your project or visit our CRISPR services page for more specific information on CRISPR-Cas9 Gene Editing services.


Applications: 

  • Recombinant protein production in CHO cells
  • Disease model in human cell lines for drug discovery
  • Gene therapy in diseased cell line
  • Target selection for drug discovery
  • High throughput preclinical screening of candidate drugs and toxicity assays
  • Custom engineering cell lines for deriving diagnostic reference standards and materials
  • Generation of a TARGATT™ master cell line by inserting an attP "docking site" for site-specific gene knock-in  (ideal model for gene function studies, reporter gene knock-in, gene overexpression, conditional/inducible gene expression)
For large DNA/ transgene insertion in cell lines, please refer to the TARGATT™ cell line editing services. The TARGATT™ integrase-based technology offers site-specific gene knock-in and is complementary to CRISPR/Cas9 technology. Both these technologies offers a broader scope for engineering physiologically predictive and advanced cell line models.  


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 10kb
  • Generate homozygous or heterozygous modified cell lines