Faithful activation of an extra‐bright red fluorescent protein in “knock‐in” Cre‐reporter mice ideally suited for lineage tracing studies
Open accessAbstract
The considerable potential of Cre recombinase as a tool for in vivo fate‐mapping studies depends on the availability of reliable reporter mice. By targeting a tandem‐dimer red fluorescent protein (tdRFP) with advanced spectral and biological properties into the ubiquitously expressed ROSA26 locus of C57BL/6‐ES cells, we have generated a novel inbred Cre‐reporter mouse with several unique characteristics. We directly demonstrate the usefulness of our reporter strain in inter‐crosses with a “universal Cre‐deleter” strain and with mice expressing Cre recombinase in a T lineage‐specific manner. Cytofluorometric and histological analyses illustrate: (i) non‐toxicity and extraordinary brightness of the fluorescent reporter, allowing quantitative detection and purification of labeled cells with highest accuracy, (ii) reliable Cre‐mediated activation of tdRFP from an antisense orientation relative to ROSA26 transcription, effectively excluding “leaky” reporter expression, (iii) absence of gene expression variegation effects, (iv) quantitative detection of tdRFP‐expressing cells even in paraformaldehyde‐fixed tissue sections, and (v) full compatibility with GFP/YFP‐based fluorescent markers in multicolor experiments. Taken together, the data show that our C57BL/6‐inbred reporter mice are ideally suited for sophisticated lineage‐tracing experiments requiring sensitive and quantitative detection/purification of live Cre‐expressing cells and their progeny.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Specific recombination in epithelial cells of the pancreas was confirmed by immunohistochemical stainings for tdRFP.
“Specific recombination in epithelial cells of the pancreas was confirmed by immunohistochemical stainings for tdRFP in Ptf1aCre/+ [35] x LSL-tdRFPKI/KI [55, 56] mice (Additional file 1: Fig. S3d) [56].”
From TBX3 is dynamically expressed in pancreatic organogenesis and fine-tunes regeneration (2023) · cites this paper as [55]
The source’s ‘Cell type-specific activation and expression of tdRFP in mice’ section reports that activated reporter cells can be monitored histologically, and Figure 8 shows tdRFP detection in spleen and lymph-node sections. It does not report Ptf1aCre crosses or confirm recombination in pancreatic epithelial cells; the citing sentence points to its own supplementary figure for that result. (LLM, conf=0.86)
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Repeat-induced gene silencing in mammals
Garrick, D., Fiering, S., Martin, D. I. K., & Whitelaw, E. (1998) · Nature Genetics · doi:10.1038/ng0198-56 · Cited by 887 · see citing works
Sauer, B. (1987) · Molecular and Cellular Biology · doi:10.1128/mcb.7.6.2087 · Cited by 482 · see citing works
Flagged first, then by citations
Research works printed but not linked: 3
- Hoess, R. H., Ziese, M. and Sternberg, N., P1 site-specific recombination: nucleotide sequence of the recombining sites. Proc. Natl. Acad. Sci. USA 1982. 79: 3398-3402.
- Campbell, R. E., Tour, O., Palmer, A. E., Steinbach, P. A., Baird, G. S., Zacharias, D. A. and Tsien, R. Y., A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. USA 2002. 99: 7877-7882.
- Hennet, T., Hagen, F. K., Tabak, L. A. and Marth, J. D., T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site- directed recombination. Proc. Natl. Acad. Sci. USA 1995. 92: 12070-12074.
Cited by
Sorted by citations
A global double-fluorescent Cre reporter mouse
Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L., & Luo, L. (2007) · genesis · doi:10.1002/dvg.20335 · Cited by 3,491 · see citing works
Fate Mapping Reveals Origins and Dynamics of Monocytes and Tissue Macrophages under Homeostasis
Yona, S., Kim, K.-W., Wolf, Y., Mildner, A., Varol, D., Breker, M., Strauss-Ayali, D., Viukov, S., Guilliams, M., Misharin, A., et al. (2013) · Immunity · doi:10.1016/j.immuni.2012.12.001 · Cited by 3,245 · see citing works
Microglia promote learning-dependent synapse formation through brain-derived neurotrophic factor
Parkhurst, C. N., Yang, G., Ninan, I., Savas, J. N., Yates, J. R., Lafaille, J. J., Hempstead, B. L., Littman, D. R., & Gan, W.-B. (2013) · Cell · doi:10.1016/j.cell.2013.11.030 · Cited by 2,599 · see citing works
Oligodendrocyte heterogeneity in the mouse juvenile and adult central nervous system
Marques, S., Zeisel, A., Codeluppi, S., van Bruggen, D., Mendanha Falcão, A., Xiao, L., Li, H., Häring, M., Hochgerner, H., Romanov, R. A., et al. (2016) · Science · doi:10.1126/science.aaf6463 · Cited by 1,246 · see citing works
Microglia remodel synapses by presynaptic trogocytosis and spine head filopodia induction
Weinhard, L., di Bartolomei, G., Bolasco, G., Machado, P., Schieber, N. L., Neniskyte, U., Exiga, M., Vadisiute, A., Raggioli, A., Schertel, A., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-03566-5 · Cited by 790 · see citing works
Fundamental properties of unperturbed haematopoiesis from stem cells in vivo
Busch, K., Klapproth, K., Barile, M., Flossdorf, M., Holland-Letz, T., Schlenner, S. M., Reth, M., Höfer, T., & Rodewald, H.-R. (2015) · Nature · doi:10.1038/nature14242 · Cited by 769 · see citing works
Lee, S., Hjerling-Leffler, J., Zagha, E., Fishell, G., & Rudy, B. (2010) · The Journal of Neuroscience · doi:10.1523/jneurosci.1869-10.2010 · Cited by 600 · see citing works
Cork, S. C., Richards, J. E., Holt, M. K., Gribble, F. M., Reimann, F., & Trapp, S. (2015) · Molecular Metabolism · doi:10.1016/j.molmet.2015.07.008 · Cited by 478 · see citing works
Richards, P., Parker, H. E., Adriaenssens, A. E., Hodgson, J. M., Cork, S. C., Trapp, S., Gribble, F. M., & Reimann, F. (2014) · Diabetes · doi:10.2337/db13-1440 · Cited by 472 · see citing works
Dulauroy, S., Di Carlo, S. E., Langa, F., Eberl, G., & Peduto, L. (2012) · Nature Medicine · doi:10.1038/nm.2848 · Cited by 418 · see citing works
Neocortical Somatostatin-Expressing GABAergic Interneurons Disinhibit the Thalamorecipient Layer 4
Xu, H., Jeong, H.-Y., Tremblay, R., & Rudy, B. (2013) · Neuron · doi:10.1016/j.neuron.2012.11.004 · Cited by 366 · see citing works
Single-cell analysis defines a pancreatic fibroblast lineage that supports anti-tumor immunity
Hutton, C., Heider, F., Blanco-Gomez, A., Banyard, A., Kononov, A., Zhang, X., Karim, S., Paulus-Hock, V., Watt, D., Steele, N., et al. (2021) · Cancer Cell · doi:10.1016/j.ccell.2021.06.017 · Cited by 337 · see citing works
Adriaenssens, A. E., Biggs, E. K., Darwish, T., Tadross, J., Sukthankar, T., Girish, M., Polex-Wolf, J., Lam, B. Y., Zvetkova, I., Pan, W., et al. (2019) · Cell Metabolism · doi:10.1016/j.cmet.2019.07.013 · Cited by 324 · see citing works
Reversible changes in pancreatic islet structure and function produced by elevated blood glucose
Brereton, M. F., Iberl, M., Shimomura, K., Zhang, Q., Adriaenssens, A. E., Proks, P., Spiliotis, I. I., Dace, W., Mattis, K. K., Ramracheya, R., et al. (2014) · Nature Communications · doi:10.1038/ncomms5639 · Cited by 259 · see citing works
Schaub, J. R., Malato, Y., Gormond, C., & Willenbring, H. (2014) · Cell Reports · doi:10.1016/j.celrep.2014.07.003 · Cited by 254 · see citing works
Broad Distribution of Hepatocyte Proliferation in Liver Homeostasis and Regeneration
Chen, F., Jimenez, R. J., Sharma, K., Luu, H. Y., Hsu, B. Y., Ravindranathan, A., Stohr, B. A., & Willenbring, H. (2020) · Cell Stem Cell · doi:10.1016/j.stem.2019.11.001 · Cited by 236 · see citing works
Adriaenssens, A. E., Svendsen, B., Lam, B. Y. H., Yeo, G. S. H., Holst, J. J., Reimann, F., & Gribble, F. M. (2016) · Diabetologia · doi:10.1007/s00125-016-4033-1 · Cited by 223 · see citing works
Neurons under T Cell Attack Coordinate Phagocyte-Mediated Synaptic Stripping
Di Liberto, G., Pantelyushin, S., Kreutzfeldt, M., Page, N., Musardo, S., Coras, R., Steinbach, K., Vincenti, I., Klimek, B., Lingner, T., et al. (2018) · Cell · doi:10.1016/j.cell.2018.07.049 · Cited by 195 · see citing works
Komuczki, J., Tuzlak, S., Friebel, E., Hartwig, T., Spath, S., Rosenstiel, P., Waisman, A., Opitz, L., Oukka, M., Schreiner, B., et al. (2019) · Immunity · doi:10.1016/j.immuni.2019.04.006 · Cited by 201 · see citing works
Kim, H., Chang, J., Shao, L., Han, L., Iyer, S., Manolagas, S. C., O'Brien, C. A., Jilka, R. L., Zhou, D., & Almeida, M. (2017) · Aging Cell · doi:10.1111/acel.12597 · Cited by 192 · see citing works
Zhan, L., Krabbe, G., Du, F., Jones, I., Reichert, M. C., Telpoukhovskaia, M., Kodama, L., Wang, C., Cho, S.-H., Sayed, F., et al. (2019) · PLOS Biology · doi:10.1371/journal.pbio.3000134 · Cited by 163 · see citing works
Activated microglia drive demyelination via <scp>CSF1R</scp> signaling
Marzan, D. E., Brügger‐Verdon, V., West, B. L., Liddelow, S., Samanta, J., & Salzer, J. L. (2021) · Glia · doi:10.1002/glia.23980 · Cited by 147 · see citing works
Senescent cells communicate via intercellular protein transfer
Biran, A., Perelmutter, M., Gal, H., Burton, D. G. A., Ovadya, Y., Vadai, E., Geiger, T., & Krizhanovsky, V. (2015) · Genes & Development · doi:10.1101/gad.259341.115 · Cited by 139 · see citing works
Van der Jeught, K., De Koker, S., Bialkowski, L., Heirman, C., Tjok Joe, P., Perche, F., Maenhout, S., Bevers, S., Broos, K., Deswarte, K., et al. (2018) · ACS Nano · doi:10.1021/acsnano.8b00966 · Cited by 139 · see citing works
Haematopoietic stem cells in perisinusoidal niches are protected from ageing
Saçma, M., Pospiech, J., Bogeska, R., de Back, W., Mallm, J.-P., Sakk, V., Soller, K., Marka, G., Vollmer, A., Karns, R., et al. (2019) · Nature Cell Biology · doi:10.1038/s41556-019-0418-y · Cited by 133 · see citing works
Treiber, N., Maity, P., Singh, K., Kohn, M., Keist, A. F., Ferchiu, F., Sante, L., Frese, S., Bloch, W., Kreppel, F., et al. (2011) · Aging Cell · doi:10.1111/j.1474-9726.2010.00658.x · Cited by 109 · see citing works
Miura, S., & Suzuki, A. (2017) · Cell Stem Cell · doi:10.1016/j.stem.2017.08.020 · Cited by 91 · see citing works
p53 in Bronchial Club Cells Facilitates Chronic Lung Inflammation by Promoting Senescence
Sagiv, A., Bar-Shai, A., Levi, N., Hatzav, M., Zada, L., Ovadya, Y., Roitman, L., Manella, G., Regev, O., Majewska, J., et al. (2018) · Cell Reports · doi:10.1016/j.celrep.2018.03.009 · Cited by 55 · see citing works
TASK-1 Potassium Channels Limit Pancreatic α-Cell Calcium Influx and Glucagon Secretion
Dadi, P. K., Luo, B., Vierra, N. C., & Jacobson, D. A. (2015) · Molecular Endocrinology · doi:10.1210/me.2014-1321 · Cited by 40 · see citing works
Thiebaud, N., Gribble, F., Reimann, F., Trapp, S., & Fadool, D. A. (2019) · Scientific Reports · doi:10.1038/s41598-019-51880-9 · Cited by 34 · see citing works
Horiguchi, M., Yoshida, M., Hirata, K., Furuyama, K., Masui, T., Uemoto, S., & Kawaguchi, Y. (2019) · FEBS Letters · doi:10.1002/1873-3468.13504 · Cited by 7 · see citing works
TBX3 is dynamically expressed in pancreatic organogenesis and fine-tunes regeneration
Melzer, M. K., Schirge, S., Gout, J., Arnold, F., Srinivasan, D., Burtscher, I., Allgöwer, C., Mulaw, M., Zengerling, F., Günes, C., et al. (2023) · BMC Biology · doi:10.1186/s12915-023-01553-x · Cited by 5 · see citing works
“Specific recombination in epithelial cells of the pancreas was confirmed by immunohistochemical stainings for tdRFP in Ptf1aCre/+ [35] x LSL-tdRFPKI/KI [55, 56] mice (Additional file 1: Fig.”
BackgroundMethodInfluential
- Authors
- Luche, H., Weber, O., Nageswara Rao, T., Blum, C., & Fehling, H. J.
- Affiliations
- Universität Ulm (DE)
- Corresponding author
- Fehling, Hans Jörg
- Venue
- European Journal of Immunology
- Publisher
- Wiley
- Published
- 2006-12-21
- Volume
- 37
- Issue
- 1
- Pages
- 43-53
- ISSN
- 0014-2980 (ISSN-L)
- 1521-4141
- OpenAlex
- W2117276705
- Cited by
- 482
- References
- 2
- Citations by year
- 2026: 15 · 2025: 21 · 2024: 17 · 2023: 20 · 2022: 19 · 2021: 42 · 2020: 40 · 2019: 29 · 2018: 34 · 2017: 24 · 2016: 35 · 2015: 34 · 2014: 31 · 2013: 35 · 2012: 21
- FWCI
- 3.81
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- European Journal of Immunology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- Cre reporter mice, Rosa26 locus, Cre-mediated recombination, lineage tracing, fate mapping, T cell lineage, fluorescence intensity, multicolor fluorescence
- MeSH terms
- Red Fluorescent Protein, Animals, Cell Line, Crosses, Genetic, Female, Gene Expression Regulation, Luminescent Proteins, Protein-Lysine 6-Oxidase, Male, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Inbred DBA, Mice, Inbred Strains, Mice, Mutant Strains, Mice, Transgenic, Extracellular Matrix Proteins, Genes, Reporter, Cell Lineage, Integrases, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
