R26R-GR: a Cre-activable dual fluorescent protein reporter mouse
You-Tzung Chen, Ming-Shian Tsai, Tsung-Lin Yang, Amy Tsu Ku, Ke-Han Huang, Cheng-Yen Huang, Fu-Ju Chou, Hsiang-Hsuan Fan, Jin-Bon Hong, Shuo-Ting Yen, Wei-Le Wang, Chang-Ching Lin, Yu-Chen Hsu, Kang-Yi Su, I-Chang Su, Chuan-Wei Jang, Richard R Behringer, Rebecca Favaro, Silvia K Nicolis, Chung-Liang Chien, Shu-Wha Lin, I-Shing Yu, You-Tzung Chen, Ming-Shian Tsai, Tsung-Lin Yang, Amy Tsu Ku, Ke-Han Huang, Cheng-Yen Huang, Fu-Ju Chou, Hsiang-Hsuan Fan, Jin-Bon Hong, Shuo-Ting Yen, Wei-Le Wang, Chang-Ching Lin, Yu-Chen Hsu, Kang-Yi Su, I-Chang Su, Chuan-Wei Jang, Richard R Behringer, Rebecca Favaro, Silvia K Nicolis, Chung-Liang Chien, Shu-Wha Lin, I-Shing Yu
Abstract
Green fluorescent protein (GFP) and its derivatives are the most widely used molecular reporters for live cell imagining. The development of organelle-specific fusion fluorescent proteins improves the labeling resolution to a higher level. Here we generate a R26 dual fluorescent protein reporter mouse, activated by Cre-mediated DNA recombination, labeling target cells with a chromatin-specific enhanced green fluorescence protein (EGFP) and a plasma membrane-anchored monomeric cherry fluorescent protein (mCherry). This dual labeling allows the visualization of mitotic events, cell shapes and intracellular vesicle behaviors. We expect this reporter mouse to have a wide application in developmental biology studies, transplantation experiments as well as cancer/stem cell lineage tracing.
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
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References
- Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263: 802–805.
- Shimomura O, Johnson FH, Saiga Y (1962) Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol 59: 223–239.
- Misgeld T, Kerschensteiner M, Bareyre FM, Burgess RW, Lichtman JW (2007) Imaging axonal transport of mitochondria in vivo. Nat Methods 4: 559–561.
- Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, et al. (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22: 1567–1572.
- Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, et al. (2008) Improving the photostability of bright monomeric orange and red fluorescent proteins. Nat Methods 5: 545–551.
- Shaner NC, Steinbach PA, Tsien RY (2005) A guide to choosing fluorescent proteins. Nat Methods 2: 905–909.
- Stewart MD, Behringer RR (2010) Confirmation of recombination site functionality in gene targeting vectors using recombinase-expressing bacteria. Methods Enzymol 477: 145–151.
- Nowotschin S, Eakin GS, Hadjantonakis AK (2009) Dual transgene strategy for live visualization of chromatin and plasma membrane dynamics in murine embryonic stem cells and embryonic tissues. Genesis 47: 330–336.
- Nowotschin S, Eakin GS, Hadjantonakis AK (2009) Live-imaging fluorescent proteins in mouse embryos: multi-dimensional, multi-spectral perspectives. Trends Biotechnol 27: 266–276.
- Stewart MD, Jang CW, Hong NW, Austin AP, Behringer RR (2009) Dual fluorescent protein reporters for studying cell behaviors in vivo. Genesis 47: 708–717.
- Livet J, Weissman TA, Kang H, Draft RW, Lu J, et al. (2007) Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature 450: 56–62.
- Snippert HJ, van der Flier LG, Sato T, van Es JH, van den Born M, et al. (2010) Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143: 134–144.
- Friedrich G, Soriano P (1991) Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev 5: 1513–1523.
- Zambrowicz BP, Imamoto A, Fiering S, Herzenberg LA, Kerr WG, et al. (1997) Disruption of overlapping transcripts in the ROSA beta geo 26 gene trap strain leads to widespread expression of beta-galactosidase in mouse embryos and hematopoietic cells. Proc Natl Acad Sci U S A 94: 3789–3794.
- Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21: 70–71.
- Mao X, Fujiwara Y, Chapdelaine A, Yang H, Orkin SH (2001) Activation of EGFP expression by Cre-mediated excision in a new ROSA26 reporter mouse strain. Blood 97: 324–326.
- Drogat B, Kalucka J, Gutierrez L, Hammad H, Goossens S, et al. (2010) Vegf regulates embryonic erythroid development through Gata1 modulation. Blood 116: 2141–2151.
- Brockschnieder D, Pechmann Y, Sonnenberg-Riethmacher E, Riethmacher D (2006) An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus. Genesis 44: 322–327.
- Srinivas S, Watanabe T, Lin CS, William CM, Tanabe Y, et al. (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 1: 4.
- Nyabi O, Naessens M, Haigh K, Gembarska A, Goossens S, et al. (2009) Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells. Nucleic Acids Res 37: e55.
- Hadjantonakis AK, Papaioannou VE (2004) Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice. BMC Biotechnol 4: 33.
- Rhee JM, Pirity MK, Lackan CS, Long JZ, Kondoh G, et al. (2006) In vivo imaging and differential localization of lipid-modified GFP-variant fusions in embryonic stem cells and mice. Genesis 44: 202–218.
- Szymczak AL, Vignali DA (2005) Development of 2A peptide-based strategies in the design of multicistronic vectors. Expert Opin Biol Ther 5: 627–638.
- Hayashi S, Lewis P, Pevny L, McMahon AP (2002) Efficient gene modulation in mouse epiblast using a Sox2Cre transgenic mouse strain. Mech Dev 119 Suppl 1: S97–S101.
- Lewandoski M, Wassarman KM, Martin GR (1997) Zp3-cre, a transgenic mouse line for the activation or inactivation of loxP-flanked target genes specifically in the female germ line. Curr Biol 7: 148–151.
- de Vries WN, Binns LT, Fancher KS, Dean J, Moore R, et al. (2000) Expression of Cre recombinase in mouse oocytes: a means to study maternal effect genes. Genesis 26: 110–112.
- Favaro R, Valotta M, Ferri AL, Latorre E, Mariani J, et al. (2009) Hippocampal development and neural stem cell maintenance require Sox2-dependent regulation of Shh. Nat Neurosci 12: 1248–1256.
- Morris RJ, Liu Y, Marles L, Yang Z, Trempus C, et al. (2004) Capturing and profiling adult hair follicle stem cells. Nat Biotechnol 22: 411–417.
- Brown DA, Rose JK (1992) Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell 68: 533–544.
- Belden WJ, Barlowe C (2001) Role of Erv29p in collecting soluble secretory proteins into ER-derived transport vesicles. Science 294: 1528–1531.
- McDowell MA, Ransom DM, Bangs JD (1998) Glycosylphosphatidylinositol-dependent secretory transport in Trypanosoma brucei. Biochem J 335 (Pt 3): 681–689.
- Sabharanjak S, Sharma P, Parton RG, Mayor S (2002) GPI-anchored proteins are delivered to recycling endosomes via a distinct cdc42-regulated, clathrin-independent pinocytic pathway. Dev Cell 2: 411–423.
- Mayor S, Riezman H (2004) Sorting GPI-anchored proteins. Nat Rev Mol Cell Biol 5: 110–120.
- Lakhan SE, Sabharanjak S, De A (2009) Endocytosis of glycosylphosphatidylinositol-anchored proteins. J Biomed Sci 16: 93.
- Meilhac SM, Adams RJ, Morris SA, Danckaert A, Le Garrec JF, et al. (2009) Active cell movements coupled to positional induction are involved in lineage segregation in the mouse blastocyst. Dev Biol 331: 210–221.
- Shioi G, Kiyonari H, Abe T, Nakao K, Fujimori T, et al. (2011) A mouse reporter line to conditionally mark nuclei and cell membranes for in vivo live-imaging. Genesis 49: 570–578.
- Abe T, Kiyonari H, Shioi G, Inoue K, Nakao K, et al. (2011) Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging. Genesis 49: 579–590.
- Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, et al. (2011) Flybow: genetic multicolor cell labeling for neural circuit analysis in Drosophila melanogaster. Nat Methods 8: 260–266.
- Hampel S, Chung P, McKellar CE, Hall D, Looger LL, et al. (2011) Drosophila Brainbow: a recombinase-based fluorescence labeling technique to subdivide neural expression patterns. Nat Methods 8: 253–259.
- Szymczak AL, Workman CJ, Wang Y, Vignali KM, Dilioglou S, et al. (2004) Correction of multi-gene deficiency in vivo using a single ‘self-cleaving’ 2A peptide-based retroviral vector. Nat Biotechnol 22: 589–594.
- Trichas G, Begbie J, Srinivas S (2008) Use of the viral 2A peptide for bicistronic expression in transgenic mice. Bmc Biology 6.
- Chen YT, Furushima K, Hou PS, Ku AT, Deng JM, et al. (2010) PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells. Stem Cells Dev 19: 763–771.
- Held M, Schmitz MH, Fischer B, Walter T, Neumann B, et al. (2010) CellCognition: time-resolved phenotype annotation in high-throughput live cell imaging. Nat Methods 7: 747–754.
- Neumann B, Walter T, Heriche JK, Bulkescher J, Erfle H, et al. (2010) Phenotypic profiling of the human genome by time-lapse microscopy reveals cell division genes. Nature 464: 721–727.
- Kondoh G, Gao XH, Nakano Y, Koike H, Yamada S, et al. (1999) Tissue-inherent fate of GPI revealed by GPI-anchored GFP transgenesis. FEBS Lett 458: 299–303.
- Nakanishi T, Ikawa M, Yamada S, Parvinen M, Baba T, et al. (1999) Real-time observation of acrosomal dispersal from mouse sperm using GFP as a marker protein. FEBS Lett 449: 277–283.
- Homans SW, Ferguson MA, Dwek RA, Rademacher TW, Anand R, et al. (1988) Complete structure of the glycosyl phosphatidylinositol membrane anchor of rat brain Thy-1 glycoprotein. Nature 333: 269–272.
- Williams AF, Gagnon J (1982) Neuronal cell Thy-1 glycoprotein: homology with immunoglobulin. Science 216: 696–703.
- Wood SA, Pascoe WS, Schmidt C, Kemler R, Evans MJ, et al. (1993) Simple and efficient production of embryonic stem cell-embryo chimeras by coculture. Proc Natl Acad Sci U S A 90: 4582–4585.
- O'Gorman S, Dagenais NA, Qian M, Marchuk Y (1997) Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. Proc Natl Acad Sci U S A 94: 14602–14607.
- Nagy A, Gertsenstein M, Vintersten K, Behringer RR (2003) Manipulating the Mouse Embryo. New York: Cold Spring Harbor Laboratory Press.
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