Fucci

Fucci

（Fluorescent ubiquitination-based cell cycle indicator）

Fucci(SA)5 and Fucci(CA)5

Two coral fluorescent proteins of distinct colors for sharp visualization of cell-cycle progression

doi: https://doi.org/10.1247/csf.23028

 

Materials

Depository Site:

Fucci – related Plasmids

Contact info: asako.sakaue-sawano’at’riken.jp

RIKEN Tsukuba
BioResource Center
Gene Engineering Division

    new!          tFucci(SA)5, tFucci(CA)5_pPBbsr2 vector

addgene

Lentiviral Vectors

pCAG-HIVgp

pCMV-VSV-G-RSV-Rev

RIKEN Tsukuba
BioResource Center

Cell lines

HeLa/Fucci
(RCB 2812)


NMuMG/Fucci
(RCB 2813)


COS/Fucci
(RCB 2814)

published in:
Cell 2008

RIKEN Tsukuba
BioResource Center
Cell Bank

HeLa/Fucci2
(RCB 2867)

NMuMG/Fucci2
(RCB 2868)

BMC CB
2011

HeLa/Fucci(CA)2
(RCB 4676)

Mol Cell
2017

HeLa/Fucci(SA)5 #16
(RCB 4917)

HeLa/Fucci(CA)5 #16
(RCB 4919)

CSF 2023

Mouse lines

FucciS/G2/M-#504
(RBRC 02706)

FucciG1-#596
(RBRC 02707)

Cell 2008

RIKEN Tsukuba
BioResource Center
Experimental Animal Division

FucciS/G2/M-#492
(RBRC 02705)

FucciG1-#639
(RBRC 02709)

PNAS 2010

FucciS/G2/M-#474
(RBRC 02704)

FucciG1-#610
(RBRC 02708)

Development
2013b

Fucci2aR
(RBRC 06511)

Cell Cycle
2014

R26p-Fucci2

R26R-Fucci2

R26-Fucci2

Development
2013a

RIKEN Kobe
Center for Developmental Biology, LARGEContact Info: mutant”at”cdb.riken.jp

Zebrafish lines

EF1α:mKO2-zCdt1(1/190)rw0405b
EF1α:mKO2-zCdt1(1/190)rw0405d

EF1α:mAG-zGem(1/100)rw0410h

EF1α:mAG-hGem(1/60)rw0412a

PNAS 2009

National BioResource Project Zebrafish

Drosophila lines

UAS-S/G2/M-Green#8 (2nd)
(DGRC# 109676)

UAS-S/G2/M-Green#18 (3rd)
(DGRC# 109677)

actp-S/G2/M-Green#3 (2nd)
(DGRC# 109678)

actp-S/G2/M-Green#12 (3rd)
(DGRC# 109679)

Mol Cell Biol 2011
Development
2011

Drosophila Genetic Resource Center

 

Information

probe Accession no. of DDBJ/EMBL/GenBank data-base
mKO2-hCdt1(30/120)      AB370332
mAG-hGeminin(1/110)      AB370333
mKO2-zCdt1(1/190)      AB505859
mAG-zGeminin(1/100)      AB505860
mAG-hGeminin(1/60)      AB505861
   
mCherry-hCdt1(30/120)      AB512478
mVenus-hGeminin(1/110)      AB512479
AmCyan-hGeminin(1/110)      AB982071
mCherry-zCdt1(1/190)      AB971706
mVenus-zGeminin(1/100)       AB971707
   
mCherry-hCdt1(1/100)Cy(-)      LC171208
mCherry-hCdt1(1/100)      LC192885
tFucci(SA)2      LC192886
tFucci(SA)2.1      LC192887
tFucci(SA)2.2      LC192888
tFucci(CA)2      LC212776
tFucci(CA)2.1      LC212777
tFucci(CA)2.2      LC212778
   
tFucci(SA)5      LC334437
tFucci(CA)5      LC334438

Fucci technology

  • Sakaue-Sawano A et al., Cell. 132: 487-498, 2008.  doi: 10.1016/j.cell.2007.12.033.
  • Sakaue-Sawano A et al., Chem. Biol. 15: 1243-1248, 2008.  doi: 10.1016/j.chembiol.2008.10.015.
  • Nishimura K et al., PLoS One. 8: e63001, 2013.  doi: 10.1371/journal.pone.0063001.
  • Oki T et al., Sci Rep. 4:4012, 2014.  doi: 10.1038/srep04012.
  • Sakaue-Sawano A & Miyawaki A: Cold Spring Harb Protoc. 2014.  doi: 10.1101/pdb.prot080408.
  • Sakaue-Sawano A et al., Mol Cell. 2017. DOI：10.1016/j.molcel.2017.10.001

This work has been selected in annual Best of Molecular Cell collection, Best of Molecular Cell 2017 http://info.cell.com/best-of-molecular-cell-tech-2017

  • Ando R, Sakaue-Sawano A, Shoda K & Miyawaki A. CSF 2023 doi: https://doi.org/10.1101/2020.03.30.015156

Fucci mouse

  • Sakaue-Sawano A et al., Cell. 132: 487-498, 2008.  doi: 10.1016/j.cell.2007.12.033.
  • Aiba Y et al., PNAS. USA. 107: 12192-12197, 2010.  doi: 10.1073/pnas.1005443107.
  • Abe T et al., Development. 140:237-246, 2013.a  doi: 10.1242/dev.084111.
  • Sakaue-Sawano A et al., Development. 140, 4624-4632, 2013.b  doi: 10.1242/dev.099226.
  • Tomura M et al., PLoS One. 8: e73801, 2013.  doi: 10.1371/journal.pone.0073801.
  • Mort RL et al., Cell Cycle. 13:17, 2681-2696, 2014.  doi: 10.4161/15384101.2015.945381.
  • Ford MJ et al., Dev Cell. 47: 4, 509-523, 2018. doi.org/10.1016/j.devcel.2018.10.027

Application for stem cell research

  • Hama H et al., Nat. Neuroscience. 14: 1481-1488, 2011.  doi: 10.1038/nn.2928.
  • Yo M et al., BBRC. 457, 7-11, 2014.  doi: 10.1016/j.bbrc.2014.12.074.

Application for drug screening

  • Sakaue-Sawano A et al., BMC Cell Biol. 12:2, 2011. doi: 10.1186/1471-2121-12-2.
  • Dan S et al., Eur. J. Cancer. 48: 936-943, 2012. doi: 10.1016/j.ejca.2011.10.006. 

Fucci zebrafish

  • Sugiyama M et al., PNAS. USA. 106: 20812-20817, 2009.  doi: 10.1073/pnas.0906464106. 
  • Fukuhara S et al., Dev Biol. 393:10-23, 2014.  doi: 10.1016/j.ydbio.2014.06.015. 

Fucci ciona

  • Ogura Y et al., Development. 138: 577-587, 2011.  doi: 10.1242/dev.053132.

Fucci drosophila

  • Nakajima Y et al., Mol. Cell Biol. 31: 2499-2512, 2011. doi: 10.1128/MCB.01046-10.
  • Makhijani K et al., Development. 138: 5379-5391, 2011.  doi: 10.1242/dev.067322. 

We are writing to relay our concern about the inaccurate terminology of the cell-cycle probe, Fucci. Whereas an increasing number of papers report cell cycle dynamics by use of the Fucci technology, the two Fucci probes, mKO2-hCdt1(30/120) and mAG-hGem(1/110), are often called mKO2-Cdt1 and mAG-Geminin, respectively. Likewise, the two Fucci2 probes are often called mCherry-Cdt1 and mVenus-Geminin. However, this terminology would mislead readers into believing that the Fucci probe contains the entire protein of Cdt1 or Geminin. In fact, more and more researchers are asking us whether such probes perturb the cell cycle regulation of recipient cells. In our original study of Fucci (Sakaue-Sawano et al. 2008), we made considerable effort to extract the regulatory domains (ubiquitination domains) from Cdt1 and Geminin; after performing long-term time-lapse imaging of numerous constructs, we demonstrated that amino acid residues 30-120 of human Cdt1 (hCdt1(30/120)) and amino acid residues 1-110 of human Geminin (hGem(1/110)) are both necessary and sufficient for the purpose. Thus, we would like to request that you use correct terminology in your forthcoming publications.

Thank you for your understanding.

Asako Sakaue-Sawano @ Laboratory for Cell Function Dynamics, CBS, RIKEN