CELL LINES
Contributor Information
- Name Jonathon Pines
- Institute University of Cambridge
Tool Details
- Tool name: Cyclin B1-Venus Reporter Cell Line [RPE1 cycB1-venus/+ KI clone 20G11]
- Alternate names: CCNB1
- Tool type: Cell Lines
- Parental cell line: RPE1
- Organism: Human
- Tissue: Eye
- Model: Reporter
- Conditional: Yes
- Description: 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.
- Research area: Cell Cycle
- 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.
- For Research Use Only
Target Details
- Target: Cyclin B1
Application Details
Handling
- Format: Frozen
- Growth medium: F12/DMEM (Sigma: D6241) + 2mM GlutaMAXĤ(Invitrogen), 10% Foetal Bovine Serum (FBS) + 0.348% sodium bicarbonate; 37oC; 5% CO2.
- Storage conditions: Liquid Nitrogen
- Shipping conditions: Dry ice
- Mycoplasma free: Yes
- Biosafety level: 1
Documentation
- Available on request
Related Tools
References
- • Collin et al., 2013. Nat Cell Biol. 15(11):1378-85. PMID: 24096242
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