Cell Line Model Generation

CRISPR Genome Editing

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CRISPR Genome Editing for Cell Lines

More than 500 unique cell line models engineered from >100 distinct parental 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 500 cell line models. We can genetically modify hard-to-transfect cells, hematopoietic/ 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    MWCL-1   BCWM-1    H929    Jurkat    

K562    KHYG-1    LAD2    MM.1s     NCI-H929     T2 cells    

TF-1     HT1080    HT29     KBM-7    KN12-Luc     LnCap    

MDA-MB231    NCI-H2228     RKO     TC32    SCC35    

SH-SY5Y    ES Cells    iPSCs  

Cancer Cell Lines Knockout
Point Mutation
Insertion
Human iPSCs Knockout
Point Mutation
Insertion
Human ES Insertion
Primate iPSCs Insertion
Human Primary Cells Knockout
Insertion
Human Fibroblasts Point Mutation
Mouse Fibroblasts Point Mutation
Rat Thyrocytes Knockout


Full service cell line editing portfolio includes:

  • Targeting vector construction and validation; cell transfection and selection; single cell cloning; expansion and cryopreservation
  • Custom edliverables: 
    • Choice of zygosity: homozygous and/or heterozygous clones
    • Choice of silent mutation at targeted site for point mutation insertion
  • Dedicated project management; detailed milestone and final reports

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

  • Knockout modifications; 
  • Gene knock-in (KI): locus-specific gene insertion; KI into safe harbor locus; gene tagging/ reporter gene insertion
  • Gene overexpression
  • Conditional / inducible gene expression; Promoter modifications
  • Gene editing/ correction, including single base changes
  • Gene replacement; gene therapy
  • Gene fusion/ translocation
  • Removal of viral sequences
  • Stable cell lines / immortalization 

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 TARGATT™ master cell lines by inserting an attP "docking site" for site-specific gene knock-in 
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.  
 
We also offer lentivirus-based stable cell line generation for difficult-to-handle cell lines: integration-free lentivirus for CRISPR-lentivirus gene knockout; broad tropism lentiviruses for efficient stable gene knock-in.

Comprehensive Technology Platforms 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

 

Support Materials

Would you like more information on our blood lineage cells editing services? Watch our webinar on CRISPR-based gene editing in blood lineage and blood-derived cell lines, presented by Dr. Jimmy Tai, Senior Scientist and Group Leader at Applied StemCell:

"CRISPR/Cas9 Editing in Blood Lineage Cells: Guidelines to enhance your CRISPR gene editing process" 

References
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