Fluorescent indicators for simultaneous reporting of all four cell cycle phases

Bryce T Bajar; Amy J Lam; Ryan K Badiee; Young-Hee Oh; Jun Chu; Xin X Zhou; Namdoo Kim; Benjamin B Kim; Mingyu Chung; Arielle L Yablonovitch; Barney F Cruz; Kanokwan Kulalert; Jacqueline J Tao; Tobias Meyer; Xiao-Dong Su; Michael Z Lin

doi:10.1038/nmeth.4045

Fluorescent indicators for simultaneous reporting of all four cell cycle phases

Nat Methods. 2016 Dec;13(12):993-996. doi: 10.1038/nmeth.4045. Epub 2016 Oct 31.

Authors

Bryce T Bajar^{1

2

3}, Amy J Lam^{1

2}, Ryan K Badiee⁴, Young-Hee Oh^{1

2

5}, Jun Chu^{1

2}, Xin X Zhou^{1

2

5}, Namdoo Kim^{1

2

5}, Benjamin B Kim^{1

2

5}, Mingyu Chung⁶, Arielle L Yablonovitch⁷, Barney F Cruz⁸, Kanokwan Kulalert⁹, Jacqueline J Tao¹, Tobias Meyer⁶, Xiao-Dong Su³, Michael Z Lin^{1

2

5}

Affiliations

¹ Department of Bioengineering, Stanford University, Stanford, California, USA.
² Department of Pediatrics, Stanford University, Stanford, California, USA.
³ School of Life Sciences, Peking University, Beijing, China.
⁴ Department of Biological Sciences, Stanford University, Stanford, California, USA.
⁵ Department of Neurobiology, Stanford University, Stanford, California, USA.
⁶ Department of Chemical and Systems Biology, Stanford University, Stanford, California, USA.
⁷ Graduate Program in Biophysics, Stanford University, Stanford, California, USA.
⁸ Department of Chemistry, Stanford University, Stanford, California, USA.
⁹ Department of Chemical Engineering, Stanford University, Stanford, California, USA.

Abstract

A robust method for simultaneous visualization of all four cell cycle phases in living cells is highly desirable. We developed an intensiometric reporter of the transition from S to G2 phase and engineered a far-red fluorescent protein, mMaroon1, to visualize chromatin condensation in mitosis. We combined these new reporters with the previously described Fucci system to create Fucci4, a set of four orthogonal fluorescent indicators that together resolve all cell cycle phases.

MeSH terms

Animals
Cell Culture Techniques
Cell Cycle / physiology*
Chromatin / metabolism
G2 Phase / physiology
HEK293 Cells
HeLa Cells
Humans
Luminescent Proteins / chemistry*
Luminescent Proteins / genetics
Mice
Mitosis
Models, Molecular
Molecular Imaging / methods*
NIH 3T3 Cells
Recombinant Fusion Proteins / chemistry*
Recombinant Fusion Proteins / genetics
Red Fluorescent Protein
S Phase / physiology
Time-Lapse Imaging / methods*

Substances

Chromatin
Luminescent Proteins
Recombinant Fusion Proteins

Abstract

MeSH terms

Substances

Grants and funding