Cells and Cell Based Assays

 

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CompoZr Cellular Reporter Cell Lines

The study of gene function has long been limited by the use of over-expression systems and by tedious staining procedures. To enable scientists to study protein function and protein localization in a live cellular system, we have generated engineered human cell lines where important proteins are fluorescently tagged at the endogenous locus through the use of CompoZr ZFN technology.

For example, CompoZr Zinc Finger Nucleases were used to fluorescently tag the α-tubulin isoform 1b (TUBA1b) in MCF10A cells. In the example, we show how we tagged the TUBA1b gene (Figure 1), the result of tagging the TUBA1b gene (Figure 2) and functional data showing the result of treating these live cells with paclitaxel (Figure 3). 

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Figure 1. Schematic of the tagged locus showing ZFN binding sites / ZFN cut site with respect to the targeted integration site. The Donor (top) has the homology arms of indicated length and the fluorescent protein (FP) sequence (green).



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Figure 2. (A) MCF10A immortalized breast epithelial knock-in cell line expressing the TUBA1B gene (α-tubulin isoform 1b) endogenously tagged with RFP at the N-terminus. Cells were imaged live in Hanks Balanced Salt solution, (Cat. No. H8264), supplemented with 2% Fetal Bovine Serum, (Cat. No. F2442), using RFP filterset (ex 530-560/em 590-650) and 40x/1.3 oil objective. (B) Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.



Bioascend. Achieve results with cell lines shaped for your research.

Figure 3. Using MCF10A RFP-TUBA1B cell line to study Paclitaxel effect on microtubules. Paclitaxel is a mitotic inhibitor used in cancer chemotherapy. Paclitaxel is thought to stabilize microtubules and as a result, interfere with the normal breakdown of microtubules during cell division. RFP tagged TUBA1B MCF10A cells were imaged by differential interference contrast (DIC) and fluorescence microscopy while applying 20 µM Paclitaxel. As time progressed, typical tubulin bundles are formed.

 

Product Offering for Cellular Reporter Cell Lines (For a shipping request outside of the USA, please contact us.)

 

Product No. Cell Line Gene Target(s) Human Gene ID Protein Function
CLL1031-1VL U2OS GFP-TUBA1B TUBA1B 10376 Globular protein - microtubule dynamics
CLL1032-1VL U2OS GFP-ACTB ACTB 60 Cytoskeletal protein - cell motility
CLL1033-1VL U2OS GFP-LMNB1 LMNB1 3912 Nuclear protein - nuclear stability & gene expression
CLL1034-1VL U2OS RFP-TUBA1B TUBA1B 10376 Globular protein - microtubule dynamics
CLL1035-1VL U2OS RFP-ACTB ACTB 60 Cytoskeletal protein - cell motility
CLL1036-1VL U2OS GFP-HMGA1 HMGA1 3159 Non-histone protein - regulation of gene expression
CLL1037-1VL U2OS GFP-ACTB RFP-TUBA1B ACTB / TUBA1B 60 / 10376 Cytoskeletal protein - cell motility / Globular protein - microtubule dynamics
CLL1038-1VL U2OS BFP-LMNB1 RFP-ACTB LMNB1 / ACTB 3912 / 60 Nuclear protein - nuclear stability & gene expression / Cytoskeletal protein - cell motility
CLL1039-1VL MCF10A RFP-TUBA1B TUBA1B 10376 Globular protein - microtubule dynamics
CLL1135-1VL SKOV3 GFP-HER2 HER2 2064 EGF receptor 2 – forms heterodimers with other EGF receptors to mediate downstream signaling pathways
CLL1136-1VL U2OS GFP-NUP98 NUP98 4928 Nucleoporin 98kD protein – protein in nuclear pore complex
CLL1139-1VL SKOV3 GFP-STAT3 STAT3 6774 Transcription factor – regulates nuclear gene expression
CLL1140-1VL A549 RFP-STAT3 STAT3 6774 Transcription factor – regulates nuclear gene expression
CLL1141-1VL A549 GFP-EGFR EGFR 1956 EGF receptor 1 – cell surface protein, binds epidermal growth factor, dimerizes and stimulates downstream signaling to promote proliferation
CLL1143-1VL SKOV3 GFP-HER2 / RFP-EGFR HER2 / EGFR 2064 / 1956 EGF receptor 2 – forms heterodimers with other EGF receptors to mediate downstream signaling pathways / EGF receptor 1 – cell surface protein, binds epidermal growth factor, dimerizes and stimulates downstream signaling to promote proliferation
CLL1149-1VL A549 GFP-CTNNB1 / RFP-LMNB1 CTNNB1 / LMNB1 1499 / 3912 Beta-catenin – protein in adherens junction, involved in regulation of cell adhesion and growth / Nuclear protein – nuclear stability & gene expression
CLL1158-1VL A549 GFP-STAT1 STAT1 6772 Transcription factor – regulates nuclear gene expression
CLL1167-1VL A549 GFP-SMAD4 SMAD4 4089 Transcription factor – regulates nuclear gene expression
CLL1218-1VL U2OS LMNB1-TUBA1B-ACTB LMNB1 / TUBA1B / ACTB 3912 / 10376 /60 Nuclear protein – nuclear stability & gene expression / Globular protein – microtubule dynamics / Cytoskeletal protein – cell motility



Resources on CompoZr Cellular Reporter Cell Lines

  • Frequently Asked Questions
  • Posters
  • Media Recommendations:
    • U2OS cell lines
      • M9309 – McCoy’s 5A Medium Modified
      • F2442 – Fetal Bovine Serum
      • C6164 – Cell Freezing Media-DMSO
    • MCF10A cell lines
      • 51448C – Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F-12
      • H1270 – Horse Serum
      • C8052 – Cholera Toxin (Vibrio cholerae)
      • I9278 – Insulin solution human (recombinant)
      • E9644 – Epidermal Growth Factor human
      • H6909 – Hydrocortisone solution
      • G3126 – L-glutamine
      • C6164 – Cell Freezing Media-DMSO
    • A549 cell lines
      • R0883 – RPMI-1640 Medium
      • F2442 – Fetal Bovine Serum
      • G7513 – L-Glutamine
      • C6164 – Cell Freezing Media-DMSO
    • SKOV cell lines
      • M8403 – McCoy's 5A Medium
      • F2442 – Fetal Bovine Serum
      • G7513 – L-Glutamine
      • C6164 – Cell Freezing Media-DMSO



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