CompoZr® Cellular Reporter Cell Lines Technical Resources and Videos

 Technical Resources

  1. John Fetter, Andrey Samsonov, Nathan Zenser, Fan Zhang, Hongyi Zhang, Dmitry Malkov. Endogenous Gene Tagging, Chapter in Methods in Molecular Biology Volume 1239, pp 231-240, 2015. 
  2. Alexandre Grassart, Aaron T. Cheng, Fan Zhang, Nathan Zenzer, Yongmei Feng, David M. Briner, Gregory D. Davis, Dmitry Malkov & David G. Drubin. Actin and dynamin2 dynamics and interplay during clathrin mediated endocytosis J. Cell Biol. Vol. 205 No. 5 721–735, 2014.
  3. Andrey Samsonov, Nathan Zenser, Fan Zhang, Hongyi Zhang, John Fetter, Dmitry Malkov. Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors.  PLoS ONE 8(7): e68391, 2013.
  4. Andrey Samsonov, Hongyi Zhang, Fan Zhang, Nathan Zenser, John Fetter, Dmitry Malkov. Novel Reporter Cell Lines For Detection Of Endogenous Pathway Activity In Live Cells. Biowire, Spring 2012, p.13-18.

 Videos

 HER2-GFP and EGFR-RFP in SKOV3 cells

SKOV3 cells that have the genomic HER2 gene endogenously tagged with GFP gene and the genomic EGFR gene - with RFP gene. The endogenous EGFR-RFP fusion protein translocates from the plasma membrane to the endosomes after stimulation with 100 ng/ml EGF while the endogenous HER2-GFP fusion protein stays at the membrane. Zinc finger nuclease (ZFN) technology was used to tag two genes (HER2 and EGFR) by inserting a fluorescent reporter sequence at the end of the coding sequence of each locus (a C-terminal fusion). DIC + epi-Fluorescence, 2 fps, 7 sec.

 

 STAT1-GFP and RFP-STAT3 in A549 cells

Endogenous STAT1/STAT3 nuclear translocation upon simultaneous activation. A549 lung carcinoma cells expressing the endogenous STAT1 protein tagged with GFP at the C-terminus and the endogenous STAT3 protein tagged with RFP at the N-terminus. Zinc finger nuclease (ZFN) technology was used to tag two genes (STAT1 and STAT3) by a fluorescent reporter sequences. The cells were preincubated with 1 µM of Hoechst 33342 and imaged live before and after addition of 100 ng/mL IFN-γ and 100 ng/mL IL-6. DIC + epi-Fluorescence, 2 fps, 9 sec.

 

 EGFR-GFP in A549 cells

A549 cells that have the genomic epidermal growth factor receptor (EGFR) gene endogenously tagged with GFP gene. The endogenous EGFR-GFP fusion protein translocates from the plasma membrane to the endosomes after stimulation with 100 ng/ml EGF. Zinc finger nuclease (ZFN) technology was used to tag EGFR gene by inserting a fluorescent reporter sequence at the end of the EGFR coding sequence (a C-terminal fusion). DIC + epi-Fluorescence, 6 fps, 16 sec.

 

 BFP-lamin B1, GFP-α-tubulin 1b, and RFP-β-actin in triple tagged U2OS cells

Zinc finger nuclease (ZFN) technology was used to tag three cytoskeletal genes by inserting a fluorescent reporter sequence behind the start codon of each locus. The integration resulted in endogenous expression of the corresponding fusion proteins. The following loci were tagged: LMNB1 (lamin B1, nuclear envelope), TUBA1B (α-tubulin 1b, microtubules), ACTB (β-actin, actin stress fibers) by BFP, GFP and RFP respectively in the same cells (U2OS). Differential interference contrast (DIC) and fluorescence microscopy images of an isolated triple knock-in clone are shown. The cells were imaged live in Hanks balanced salt solution supplemented with 2% fetal bovine serum using corresponding filter sets and 40x/1.3 oil objective. The bar is 25 μm. 7 fps, 5 sec.

 

 Paclitaxel effect on RFP-α-tubulin in MCF10A cells

Paclitaxel (or Taxol -- cancer drug) effect on microtubules. MCF10A cells with a cytoskeletal protein α-tubulin RFP tagged. Zinc finger nuclease (ZFN) technology was used to tag TUBA1B gene by inserting a fluorescent reporter sequence behind the start codon of this locus. DIC + epi-Fluorescence, 10 fps, 22 sec.

 

 GFP-lamin-B1 in U2OS - cell division

U2OS cell division. A protein that forms the inner nuclear lining, lamin B1, is GFP tagged. Zinc finger nuclease (ZFN) technology was used to tag LMNB1 gene by inserting a fluorescent reporter sequence behind the start codon of this locus. DIC + epi-Fluorescence, 10 fps, 18 sec.

 

 GFP-based biosensor detects endogenous EGF receptor activity in A549 cells

Detection of endogenous EGFR phosphorylation and internalization by a GFP-based biosensor. EGF causes the biosensor to translocate towards the plasma membrane and bind EGFR followed by internalization in A549 cells. The nucleus was labeled with DRAQ5. DIC + epi-Fluorescence, 6 fps, 12 sec.

 

 GFP-β-actin in U2OS cells

Lamellipodia formation by actin polymerization. U2OS cells with a cytoskeletal protein β-actin GFP tagged. Zinc finger nuclease (ZFN) technology was used to tag ACTB gene by inserting a fluorescent reporter sequence behind the start codon of this locus. DIC + epi-Fluorescence, 10 fps, 19 sec.

 

 HMGA1-GFP in U2OS - cell division

U2OS cell division. A non-histone DNA binding protein HMGA1 is GFP tagged. Zinc finger nuclease (ZFN) technology was used to tag HMGA1 gene by inserting a fluorescent reporter sequence in front of the stop codon of this locus. DIC + epi-Fluorescence, 6fps, 19 sec.

 

 GFP-lamin-B1 and RFP-β-actin

Actin supported ruffles. Dual tagged U2OS cells: GFP-lamin B1 (nuclear envelope), RFP-β-actin (actin stress fibers). Zinc finger nuclease (ZFN) technology was used to tag two genes (LMNB1 and ACTB) by inserting a fluorescent reporter sequence behind the start codon of each locus. DIC + epi-Fluorescence, 10fps, 17 sec.