• Cell Line Model Generation

Cell Line Models

Applied StemCell’s CRISPR/Cas9 cell line modeling experts have successfully engineered more than 500 distinct cell line models in > 100 distinct mammalian cell lines from different species. We use highly optimized and efficient CRISPR/Cas9 genome editing strategies and protocols to engineer or correct mutations in a variety of cell lines such as cancer cells, hard-to-transfect cells, blood lineage cells, stem cells and many more with a > 97% success rate.

Our full service cell line editing portfolio includes:

  • Targeting vector construction and validation; cell transfection and selection; single cell cloning; expansion and cryopreservation

  • Custom deliverables: choice of homozygous and/or heterozygous clones; point mutation with/without silent mutation

Cell Line Model Generation Categories

CRISPR/Cas9
Cell Line
Service

Our CRISPR/Cas9 service uses proprietary gRNA design and protocols to genetically modify any gene of interest in hundreds of mammalian cell lines.

CRISPR/Cas9
Cell Line
Service

CRISPR/Cas9 Cell Line
Service - Hematopoietic Cells
(Jurkat and TF-1)

Applied StemCell has optimized the CRISPR/Cas9 technology to achieve the highest success rate for genome editing of human blood lineage cell lines.

CRISPR/Cas9 Cell Line
Service - Hematopoietic Cells
(Jurkat and TF-1)

CRISPR/Cas9
Genome Editing in
Stem Cells

CRISPR/Cas9 genome editing in control/ patient iPSCs to engineer or correct mutations for predictive in vitro modeling of human biology and disease.

CRISPR/Cas9 Genome
Editing in
Stem Cells

Publications

Applied StemCell publications and citations:

  • Selvan, N., George, S., Serajee, F. J., Shaw, M., Hobson, L., Kalscheuer, V. M., ... & Schwartz, C. E. (2018). O-GlcNAc transferase missense mutations linked to X-linked intellectual disability deregulate genes involved in cell fate determination and signaling. Journal of Biological Chemistry, jbc-RA118.

  • Smalley, E. (2018). FDA warns public of dangers of DIY gene therapy. https://doi.org/10.1038/nbt0218-119

  • Chai, S., Wan, X., Ramirez-Navarro, A., Tesar, P. J., Kaufman, E. S., Ficker, E., ... & Deschênes, I. (2018). Physiological genomics identifies genetic modifiers of long QT syndrome type 2 severity. The Journal of clinical investigation, 128(3).

  • Boi, S., Ferrell, M. E., Zhao, M., Hasenkrug, K. J., & Evans, L. H. (2018). Mouse APOBEC3 expression in NIH 3T3 cells mediates hypermutation of AKV murine leukemia virus. Virology, 518, 377-384. https://doi.org/10.1016/j.virol.2018.03.014.

  • Molinski, S. V., et al. (2017). Orkambi® and amplifier co‐therapy improves function from a rare CFTR mutation in gene‐edited cells and patient tissue. EMBO Molecular Medicine, e201607137.

  • Petrovic, P. B. (2017). Myosin Phosphatase Rho-interacting Protein Regulates DDR1-mediated Collagen Tractional Remodeling (Doctoral dissertation, University of Toronto (Canada)).

  • Peng, L., Zhang, H., Hao, Y., Xu, F., Yang, J., Zhang, R., ... & Chen, C. (2016). Reprogramming macrophage orientation by microRNA 146b targeting transcription factor IRF5. EBioMedicine, 14, 83-96.

  • Hu, J. K., Crampton, J. C., Locci, M., & Crotty, S. (2016). CRISPR-mediated Slamf1Δ/Δ Slamf5Δ/Δ Slamf6Δ/Δ triple gene disruption reveals NKT cell defects but not T follicular helper cell defects. PloS one, 11(5), e0156074.

  • Smalley, E. (2016). CRISPR mouse model boom, rat model renaissance. Nature Biotechnology. 34, 893–894.

  • Baker, M. (2014). Gene editing at CRISPR speed. Nature biotechnology, 32(4), 309-313.

CRISPR Technology:

Technical Details

More than 500 unique cell line models engineered from >100 distinct parental cell lines!

Cell Type Modification Type
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

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

Partial list of cell line engineering projects completed by Applied StemCell:

  • 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 Genome 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
Have Questions?

An Applied StemCell technical expert is happy to help, contact us today!