Cell cycle tracker stocks

P{UAS-RGB.cct} has the following structure. 

This image shows the structure of the P{UAS-RGB.cct} transgenic construct

As shown in the figure below, it expresses:

• EBFP2 with a nuclear localization signal fused to a degron sequence from dup/Cdt1 for degradation during S phase.

• tdTomato nuclear localization signals fused to degron sequences from CycB for degradation during late mitosis and early G1 phase.

• EGFP fused to PCNA to give a unique subnuclear localization during S phase.

The panels in this figure show the expression of EBFP2 under control of Cdt1 regulatory sequences, the expression of tdTomato under the control of CycB regulatory sequences and the expression of EGFP under the control of PCNA regulatory sequences from the UAS-RGB.cct construct in the same cells during different cell cycle phases (G1, early S, late S, early G2, G2, late G2 and M)

It was described in Handke et al. (2014).” Towards long term cultivation of Drosophila wing imaginal discs in vitro” PLoS ONE 9: e107333.

P{UAS-RB.cct} [No FBid yet] is identical except for omission of the T2A-EGFP-PCNA sequence.

These constructs provide alternatives to the fluorescent ubiquitination-based cell cycle indicator (Fly-FUCCI) for marking cell cycle stages.


Related links   Fly-FUCCI   UAS   GAL4   QUAS   Fluors