MAPK, KRAS, BRAF Pathway Isogenic Cell Lines

About the MAPK Pathway Isogenic Cell Lines

The MAPK signaling pathway encompasses a series of signal transduction events that begin with the engagement of EGFR at the outer cell membrane, and proceed through KRAS, BRAF, MEK, and ERK.  This signaling cascade ultimately results in transcriptional activation of key genes that promote cellular proliferation, survival, differentiation, motility, and angiogenesis1.  As such, the MAPK pathway is one of the most frequently activated pathways in cancer2, and several drugs have pharmacogenomics profiles that depend upon the MAPK mutational status3.

Applied StemCell has engineered a series of cell lines that feature diverse mutations in EGFR, KRAS, and BRAF.  These cell lines provide an ideal toolset for enhancing our understanding of cancer biology, or for high throughput screening to identify lead compounds for cancer therapeutics.

mapk-figure

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  EGFR Mutations Covered by ASC
  CDS Mutation AA Mutation Mutation ID Mutation State
Exon 18 c.2155G>A p.G719A COSM6252  
c.2155G>T p.G719C COSM6253 Homozygous
c.2156G>C p.G719A COSM6239 Homozygous
Exon 19 c.2239_2256del18 p.L747_S752delLREATS   Homozygous
c.2240_2257del18 p.L747_P753>S   Homozygous
c.2238_2255del18  p.E746_S752>D   Homozygous
c.2239_2258>CA p.L747_P753>Q   Homozygous
c.2237_2254del18 p.E746_S752>A   Homozygous
c.2237_2255>T p.E746_S752>V   Homozygous
c.2236_2253del18 p.E746_T751delELREAT   Homozygous
c.2237_2251del15 p.E746_T751>A   Homozygous
c.2239_2253del15 p.L747_T751delLREAT   Homozygous
c.2240_2254del15 p.L747_T751delLREAT   Homozygous
c.2237_2252>T p.E746_T751>V   Homozygous
c.2235_2252>AAT p.E746_T751>I   Homozygous
c.2233_2247del15 p.K745_E749delKELRE   Homozygous
c.2235_2249del15 p.E746_A750delELREA   Homozygous
c.2235_2250del15 p.E746_A750delELREA   Homozygous
c.2239_2251>C p.L747_T751>P   Homozygous
c.2239_2251>C p.L747_T751>S   Homozygous
c.2238_2252>GCA p.L747_T751>Q   Homozygous
c.2239_2248TTAAGAGAAG>C p.L747_A750>P   Homozygous
c.2238_2248>GC p.L747_A750>P   Homozygous
c.2239_2247del9 p.L747_E749delLRE   Homozygous
c.2235_2248>AATTC p.E746_A750>IP   Homozygous
Exon 20 c.2303G>T p.S768I   Homozygous
c.2307_2308insGCCAGCGTG p.V769_D770insASV   Homozygous
c.2310_2311insGGT p.D770_N771insG   Homozygous
c.2319_2320insCAC p.H773_V774insH   Homozygous
c.2369C>T p.T790M   Homozygous
Exon 21 c.2573T>G p.L858R    Homozygous
c.2582T>A p.L861Q    Homozygous

 

  KRAS Mutations Covered by ASC
  CDS Mutation AA Mutation Mutation ID Mutation State
Exon 2 c.34G>T  p.G12C  COSM516   
c.34G>A p.G12S COSM517  
c.34G>C p.G12R COSM518  
c.35G>T p.G12V COSM520 Homozygous
c.35G>A p.G12D COSM521  
c.35G>C p.G12A COSM522  
c.37G>T  p.G13C COSM527 Homozygous
c.37G>A p.G13S COSM528 Homozygous
c.37G>C p.G13R COSM529 Homozygous
c.38G>A p.G13D COSM532 Homozygous
c.38G>C p.G13A COSM533  
c.38G>T p.G13V COSM534 Homozygous
Exon 15 c.1798_1799GT>AA p.V600K COSM473 Homozygous
c.1798_1799GT>AG p.V600R COSM474 Homozygous
c.1799_1800TG>AA p.V600E COSM475 Homozygous 
c.1799T>A p.V600E COSM476 Homozygous
c.1799_1800TG>AT p.V600D COSM477 Homozygous
c.1798G>A p.V600M COSM1130 Homozygous
c.1799T>G p.V600G COSM6147 Homozygous
Technical Details

Engineering Clinically Relevant Mutations - Control every aspect of your workflow

- Footprint-free mutations

- Homozygous mutations

- Paired isogenic mutation and wild-type cell lines

TECHNICAL-MAPK-Isogenic-cellline-engineering

Support Materials

ONCOREF™ Reference Standards: Application of CRISPR/Cas9 to the Generation of Isogenic Cell Lines and Reference Materials (October 2016)

2016-Oncoref-webinar

 

 

 

 

 

 

 

 

 

 

CRISPR/Cas9 is rapidly enabling the development of new tools for enhancing our understanding oncogenic mutations in cancer. In order to aid in advancing cancer diagnosis and treatment, Applied StemCell has recently engineered a series of 40 isogenic cell lines that feature diverse mutations in the MAPK pathway. These mutant lines are available as isogenic pairs for applications in lead compound discovery, or as FFPE and nucleic acid reference materials for assay development. This webinar will focus on ASC’s efforts in developing these research tools, as well as applications of the materials for the advancement of cancer research.

Highlights of this talk:

  • Overview of molecular reference materials 
  • Workflow and QC for ONCOREF™ cell line generation
  • Advantages of CRISPR-engineered molecular reference standards
  • Applications of reference materials in assay development
  • Q & A
FAQ

How are your cell lines generated?

 Applied StemCell uses CRISPR/Cas9 technology for cell line engineering.  This technology allows for footprint-free gene modification, which means that there are no selection markers or other genomic footprints that may impact your scientific results.

How do you validate mutational status in your cell lines?

All of our MAPK mutation panel cell lines are expanded from single cell clones, and the targeted mutations are validated at the genomic level by Sanger sequencing.  By generating clonal populations of mutant cell lines, we can ensure the highest quality and homogeneity of the engineered cells.

What if I need a cell line that you don’t currently offer?

In addition to providing off-the-shelf cell lines, ASC offers comprehensive cell line engineering services.  Just tell us your gene of interest and desired mutation, and we will design and engineer the cell line for you.  Talk to one of our knowledgeable sales representatives for more details.

References
  1. Akinleye, A., Furqan, M., Mukhi, N., Ravella, P. & Liu, D. MEK and the inhibitors: from bench to bedside. J. Hematol. Oncol. 6, 1–11 (2013).
  2. Santarpia, L., Lippman, S. M. & El-Naggar, A. K. Targeting the MAPK–RAS–RAF signaling pathway in cancer therapy. Expert Opin. Ther. Targets 16, 103–119 (2012).
  3. Relling, M. V & Evans, W. E. Pharmacogenomics in the clinic. Nature 526, 343–350 (2015).
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