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Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice

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Fucci2a : a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. / Mort, Richard Lester; Ford, Matthew Jonathan; Sakaue-Sawano, Asako; Lindstrom, Nils Olof; Casadio, Angela; Douglas, Adam Thomas; Keighren, Margaret Anne; Hohenstein, Peter; Miyawaki, Atsushi; Jackson, Ian James.

In: Cell Cycle (Georgetown, Tex.), Vol. 13, No. 17, 2014, p. 2681-2696.

Research output: Contribution to journalJournal article

Harvard

Mort, RL, Ford, MJ, Sakaue-Sawano, A, Lindstrom, NO, Casadio, A, Douglas, AT, Keighren, MA, Hohenstein, P, Miyawaki, A & Jackson, IJ 2014, 'Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice', Cell Cycle (Georgetown, Tex.), vol. 13, no. 17, pp. 2681-2696. https://doi.org/10.4161/15384101.2015.945381

APA

Mort, R. L., Ford, M. J., Sakaue-Sawano, A., Lindstrom, N. O., Casadio, A., Douglas, A. T., Keighren, M. A., Hohenstein, P., Miyawaki, A., & Jackson, I. J. (2014). Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. Cell Cycle (Georgetown, Tex.), 13(17), 2681-2696. https://doi.org/10.4161/15384101.2015.945381

Vancouver

Mort RL, Ford MJ, Sakaue-Sawano A, Lindstrom NO, Casadio A, Douglas AT et al. Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. Cell Cycle (Georgetown, Tex.). 2014;13(17):2681-2696. https://doi.org/10.4161/15384101.2015.945381

Author

Mort, Richard Lester ; Ford, Matthew Jonathan ; Sakaue-Sawano, Asako ; Lindstrom, Nils Olof ; Casadio, Angela ; Douglas, Adam Thomas ; Keighren, Margaret Anne ; Hohenstein, Peter ; Miyawaki, Atsushi ; Jackson, Ian James. / Fucci2a : a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. In: Cell Cycle (Georgetown, Tex.). 2014 ; Vol. 13, No. 17. pp. 2681-2696.

Bibtex

@article{05973bc183b64962ac245de0142b4684,
title = "Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice",
abstract = "Markers of cell cycle stage allow estimation of cell cycle dynamics in cell culture and during embryonic development. The Fucci system incorporates genetically encoded probes that highlight G1 and S/G2/M phases of the cell cycle allowing live imaging. However the available mouse models that incorporate Fucci are beset by problems with transgene inactivation, varying expression level, lack of conditional potential and/or the need to maintain separate transgenes-there is no transgenic mouse model that solves all these problems. To address these shortfalls we re-engineered the Fucci system to create 2 bicistronic Fucci variants incorporating both probes fused using the Thosea asigna virus 2A (T2A) self cleaving peptide. We characterize these variants in stable 3T3 cell lines. One of the variants (termed Fucci2a) faithfully recapitulated the nuclear localization and cell cycle stage specific florescence of the original Fucci system. We go on to develop a conditional mouse allele (R26Fucci2aR) carefully designed for high, inducible, ubiquitous expression allowing investigation of cell cycle status in single cell lineages within the developing embryo. We demonstrate the utility of R26Fucci2aR for live imaging by using high resolution confocal microscopy of ex vivo lung, kidney and neural crest development. Using our 3T3 system we describe and validate a method to estimate cell cycle times from relatively short time-lapse sequences that we then apply to our neural crest data. The Fucci2a system and the R26Fucci2aR mouse model are compelling new tools for the investigation of cell cycle dynamics in cell culture and during mouse embryonic development.",
keywords = "3T3 Cells, Animals, Cell Cycle, Cell Proliferation, Cell Survival, Embryo, Mammalian, Embryonic Stem Cells, G1 Phase, Gene Expression, Genes, Reporter, Humans, Integrases, Kidney, Luminescent Proteins, Lung, Mice, Mitosis, Morphogenesis, Organ Specificity, Time Factors, Time-Lapse Imaging, Journal Article, Research Support, Non-U.S. Gov't",
author = "Mort, {Richard Lester} and Ford, {Matthew Jonathan} and Asako Sakaue-Sawano and Lindstrom, {Nils Olof} and Angela Casadio and Douglas, {Adam Thomas} and Keighren, {Margaret Anne} and Peter Hohenstein and Atsushi Miyawaki and Jackson, {Ian James}",
year = "2014",
doi = "10.4161/15384101.2015.945381",
language = "English",
volume = "13",
pages = "2681--2696",
journal = "Cell Cycle (Georgetown, Tex.)",
issn = "1538-4101",
publisher = "Landes Bioscience",
number = "17",

}

RIS

TY - JOUR

T1 - Fucci2a

T2 - a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice

AU - Mort, Richard Lester

AU - Ford, Matthew Jonathan

AU - Sakaue-Sawano, Asako

AU - Lindstrom, Nils Olof

AU - Casadio, Angela

AU - Douglas, Adam Thomas

AU - Keighren, Margaret Anne

AU - Hohenstein, Peter

AU - Miyawaki, Atsushi

AU - Jackson, Ian James

PY - 2014

Y1 - 2014

N2 - Markers of cell cycle stage allow estimation of cell cycle dynamics in cell culture and during embryonic development. The Fucci system incorporates genetically encoded probes that highlight G1 and S/G2/M phases of the cell cycle allowing live imaging. However the available mouse models that incorporate Fucci are beset by problems with transgene inactivation, varying expression level, lack of conditional potential and/or the need to maintain separate transgenes-there is no transgenic mouse model that solves all these problems. To address these shortfalls we re-engineered the Fucci system to create 2 bicistronic Fucci variants incorporating both probes fused using the Thosea asigna virus 2A (T2A) self cleaving peptide. We characterize these variants in stable 3T3 cell lines. One of the variants (termed Fucci2a) faithfully recapitulated the nuclear localization and cell cycle stage specific florescence of the original Fucci system. We go on to develop a conditional mouse allele (R26Fucci2aR) carefully designed for high, inducible, ubiquitous expression allowing investigation of cell cycle status in single cell lineages within the developing embryo. We demonstrate the utility of R26Fucci2aR for live imaging by using high resolution confocal microscopy of ex vivo lung, kidney and neural crest development. Using our 3T3 system we describe and validate a method to estimate cell cycle times from relatively short time-lapse sequences that we then apply to our neural crest data. The Fucci2a system and the R26Fucci2aR mouse model are compelling new tools for the investigation of cell cycle dynamics in cell culture and during mouse embryonic development.

AB - Markers of cell cycle stage allow estimation of cell cycle dynamics in cell culture and during embryonic development. The Fucci system incorporates genetically encoded probes that highlight G1 and S/G2/M phases of the cell cycle allowing live imaging. However the available mouse models that incorporate Fucci are beset by problems with transgene inactivation, varying expression level, lack of conditional potential and/or the need to maintain separate transgenes-there is no transgenic mouse model that solves all these problems. To address these shortfalls we re-engineered the Fucci system to create 2 bicistronic Fucci variants incorporating both probes fused using the Thosea asigna virus 2A (T2A) self cleaving peptide. We characterize these variants in stable 3T3 cell lines. One of the variants (termed Fucci2a) faithfully recapitulated the nuclear localization and cell cycle stage specific florescence of the original Fucci system. We go on to develop a conditional mouse allele (R26Fucci2aR) carefully designed for high, inducible, ubiquitous expression allowing investigation of cell cycle status in single cell lineages within the developing embryo. We demonstrate the utility of R26Fucci2aR for live imaging by using high resolution confocal microscopy of ex vivo lung, kidney and neural crest development. Using our 3T3 system we describe and validate a method to estimate cell cycle times from relatively short time-lapse sequences that we then apply to our neural crest data. The Fucci2a system and the R26Fucci2aR mouse model are compelling new tools for the investigation of cell cycle dynamics in cell culture and during mouse embryonic development.

KW - 3T3 Cells

KW - Animals

KW - Cell Cycle

KW - Cell Proliferation

KW - Cell Survival

KW - Embryo, Mammalian

KW - Embryonic Stem Cells

KW - G1 Phase

KW - Gene Expression

KW - Genes, Reporter

KW - Humans

KW - Integrases

KW - Kidney

KW - Luminescent Proteins

KW - Lung

KW - Mice

KW - Mitosis

KW - Morphogenesis

KW - Organ Specificity

KW - Time Factors

KW - Time-Lapse Imaging

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.4161/15384101.2015.945381

DO - 10.4161/15384101.2015.945381

M3 - Journal article

C2 - 25486356

VL - 13

SP - 2681

EP - 2696

JO - Cell Cycle (Georgetown, Tex.)

JF - Cell Cycle (Georgetown, Tex.)

SN - 1538-4101

IS - 17

ER -