• Cancer Research

Cancer Research

We are experts in cell line gene editing! ASC supports cancers researchers by providing advanced, physiologically relevant cell line models with precise genetic modifications. Our multi-approach designing strategy and combination of technologies such as in-licensed CRISPR/Cas9, proprietary TARGATT™, and lentiviral technologies enables us to engineer your perfect stable cell line model. We can genetically engineer a variety of cell lines (see Technical Details section) with customizable deliverables. Also, explore our series of genetically modified cell lines/ isogenic cell lines for Cas9 expressing cell lines and iPSC-reporter master cells lines that are ideal for generating your own isogenic cancer disease cell lines models.

Publications

Cell Line Models:  Knock-Out, Knock-in, Point Mutation

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 investigation128(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. Virology518, 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. EBioMedicine14, 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 one11(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 biotechnology32(4), 309-313.

CRISPR Technology:

Technical Details

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
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

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  

Customizable deliverables:

  • Footprint-free genome editing
  • Custom heterozygous/ homozygous clones
  • And point mutations without silent mutations

A partial list of genetically modified cell line models:

  • Gene knockout (KO): frame shift; fragment excision, stop cassette insertion, double KO
  • Gene knock-in (KI): point mutation, reporter gene, small and large fragment insertion (locus-specific/ safe harbor locus)
  • Controlled gene expression models: gene overexpression; conditional/ inducible gene expression; promoter modifications
  • Gene editing/ correction, including single base changes/ point mutation
  • Gene replacement; gene therapy
  • Gene fusion/ translocation
  • Removal of viral sequences
  • Master cell line generation

Don’t see a particular model you are interested in? Contact us to learn about the full scope of our expertise and get a cell line model engineered precisely to your project requirements.

Applications:

  • Modeling cancer disease for understanding genetic role in cancer progression
  • Pharmacogenomic studies
  • Drug efficacy and toxicity screening
  • Drug combination studies
  • Antibody validation

Choosing the right genome editing technology:
Applied StemCell uses two complementary genome editing technologies to generate advanced cell line and animal models very efficiently and effectively: the CRISPR/Cas9 technology and our propriety site-specific gene integration technology, TARGATT™ for large fragment (up to 20 kb) knock-in into a safe harbor locus.

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) 

Have Questions?

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