CELL LINES
Contributor Information
- Name Jonathon Pines
- Institute University of Cambridge
Tool Details
- Tool name: Cyclin A2-Venus Reporter Cell Line [RPE1 cycA2-venus/+ KI clone D6]
- Alternate names: CCNA2
- Tool type: Cell Lines
- Parental cell line: RPE1
- Organism: Human
- Tissue: Eye
- Model: Reporter
- Conditional: No
- Description: Somatic knock-in reporter cell line with endogenous expression of Cyclin A2 fused to the yellow fluorescent protein Venus (Cyclin A2-Venus fusion protein). Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. Cyclin A binds and activates CDC2 or CDK2 kinases, and thus this cell line functions as a reporter for both cell cycle G1/S and G2/M transitions. See also the Mad2 and Cyclin B1 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 A2 levels and activity.
- Research area: Cell biology
- Production details: To generate the cell line recombinant adenovirus-associated virus (rAAV)-mediated gene targeting was used to introduce the open reading frame (ORF) of yellow fluorescent protein (Venus) into the last exon of one allele of the CCNA2 (Cyclin A2) 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 A2 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 A2
Application Details
Handling
- Format: Frozen
- Growth medium: F12/DMEM (Sigma: D6241) + 2mM GlutaMAX?��¤(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
Related Tools
References
- • Collin et al., 2013. Nat Cell Biol. 15(11):1378-85. PMID: 24096242
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