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Cyclin B1-Venus Reporter Cell Line [RPE1 cycB1-venus/+ KI clone 20G11]

Invented by Dr Jonathon Pines at Gurdon Institute

Info

Catalogue Number 153464
Antigen/Gene or Protein Targets Cyclin B1
Parental Line RPE1
Synonyms CCNB1
Host Human
Tissue Eye
Model Reporter
Relevance Somatic knock-in reporter cell with endogenous expression of cyclin B1 fused to the yellow fluorescent protein Venus (cyclin B1-Venus fusion protein). Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This Cyclin B1 cell line functions as a cell cycle reporter across the G2/mitotic-specific phase of the cell cycle. See also the Mad2 and Cyclin A2 versions of these cell lines for a broader analysis of the different phases of the cell cycle. This cell line is an endogenous reporter for cyclin B1 levels and activity.
Production Details To generate the cell line recombinant adenovirus-associated virus (rAAV)-mediated gene targeting was used to fuse the open reading frame (ORF) of yellow fluorescent protein (Venus) with exon 9 of the CCNB1 (Cyclin B1) gene in hTert-RPE1 cells (RPE1; retinal pigment epithelial). RPE1 cells were chosen because they have a normal diploid karyotype, are not transformed and exhibit little cell death when arrested in mitosis; tagging the endogenous Cyclin B1 protein in RPE1 cells avoided the complications of mosaic protein levels in the cell population produced by ectopic expression.
Conditional No
Research Area Cell Cycle
Recommended Growing Conditions F12/DMEM (Sigma: D6241) + 2mM GlutaMAX(Invitrogen), 10% Foetal Bovine Serum (FBS) + 0.348% sodium bicarbonate; 37oC; 5% CO2.

References

There are 2 reference entries for this reagent.

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References: 2 entries

Collin et al. 2013. Nat Cell Biol. 15(11):1378-85. PMID: 24096242.

The spindle assembly checkpoint works like a rheostat rather than a toggle switch.

Europe PMC ID: 24096242


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References: 2 entries

Collin et al. 2013. Nat Cell Biol. 15(11):1378-85. PMID: 24096242.

The spindle assembly checkpoint works like a rheostat rather than a toggle switch.


Add a reference