Sigma® Life Science has applied the revolutionary CompoZr Zinc Finger Nuclease technology to create an unparalleled range of genetically modified mammalian cell lines for use in areas such as basic research, target validation, drug discovery and drug development.
CompoZr Oncology Disease Model Cell Lines
An individual patient’s response to therapy may vary depending on their unique genotype. To better understand the genetics of cancer, we have generated genetically-defined mutations in human cell lines that model patient-relevant genome alterations. Our oncology offerings focus on colorectal carcinoma using the human DLD1 and SW48 cell lines and on lung cancer using the human A549 cell line. These tools will enable researchers to study disease gene targets in an isogenic setting, under the endogenous promoter and enable better avenues for therapeutic research and drug screening.
Knockout Cell Lines Available
Cell Culture Media Components
DLD1 cell lines
A549 cell lines
SW48 cell lines
Advantages and Benefits
To generate knockout mutations of an endogenous gene, a pair of ZFNs were designed and transduced into the cells to make a specific double-strand break (DSB) in the coding region of the gene of interest. During the process of non-homologus end joining (NHEJ) to repair the DSB, imperfect repair results in cells containing mutations (insertions or deletions of DNA) that result in nonsense transcripts targeted for degradation, yielding a gene knockout phenotype.
Figure 1. (A) Each Zinc Finger Nuclease (ZFN) consists of two functional domains: A DNA-binding domain comprised of a chain of zinc finger modules, each recognizing a unique triplet (3 bp) sequence of DNA. Four to six zinc finger modules are stitched together to form a Zinc Finger Protein (ZFP), with specificity of ≥12 bp. A DNA-cleaving domain comprised of the nuclease domain of FokI is attached to the ZFPs. When the DNA-binding and DNA-cleaving domains are fused together, a highly specific pair of 'genomic scissors' is created that binds with 24-36 bp specificity of the ZFPs and cleaves the DNA. (B) The addition of zinc finger nucleases to the cell results in creation of a double-strand break at the target site. This double-strand break is repaired by one of two endogenous repair pathways, either the non-homologous end joining (NHEJ) or the homologous recombination (HR) pathway. NHEJ is used to create gene knockouts while HR is utilized for targeted integration.
Example of BAX Gene Knockout in all Four Alleles in A549 Lung Carcinoma Cells
Many cell immortalized cell lines (e.g. A549) exhibit polyploidy of gene alleles and the ZFN technology can be use to knockout a specific gene in all alleles of the cell.
Figure 2. Knockout of tetraploid BAX in A549 cells using ZFNs.
Where to use CompoZr Disease Model Cell Lines