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Perinatal induction of Cre recombination with tamoxifen.

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Lizen, Benoît [UCL] Claus, Mélissa [UCL] Jeannotte, Lucie [Université Laval, Québec, Canada] Rijli , F.M. [Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland] Gofflot, Françoise [UCL]

Temporal control of site-specific recombination is commonly achieved by using a tamoxifen-inducible form of Cre or Flp recombinases. Although powerful protocols of induction have been developed for gene inactivation at adult stages or during embryonic development, induction of recombination at late gestational or early postnatal stages is still difficult to achieve. In this context, using the ubiquitous CMV-CreERT2 transgenic mice, we have tested and validated two procedures to achieve recombination just before and just after birth. The efficiency of recombination was evaluated in the brain, which is known to be more problematic to target. For the late gestation treatment with tamoxifen, different protocols of complementary administration of progesterone and estrogen were tested. However, delayed delivery and/or mortality of pups due to difficult delivery were always observed. To circumvent this problem, pups were collected from tamoxifen-treated pregnant dams by caesarian section at E18.5 and given to foster mothers. For postnatal treatment, different dosages of tamoxifen were administered by intragastric injection to the pups during 3 or 4 days after birth. The efficiency of these treatments was analyzed at P7 using a transgenic reporter line. They were also validated with the Hoxa5 conditional allele. In conclusion, we have developed efficient procedures that allow achieving efficient recombination of floxed alleles at perinatal stages. These protocols will allow investigating the late/adult functions of many developmental genes, whose characterization has been so far restricted to embryonic development.

Document type Article de périodique (Journal article) – Article de recherche
Access type Accès restreint
Publication date 2015
Language Anglais
Journal information "Transgenic Research" - Vol. 24, p. 1065-1077 (2015)
Peer reviewed yes
Publisher Springer Netherlands ((Netherlands) Dordrecht)
issn 0962-8819
e-issn 1573-9368
Publication status Publié
Affiliations UCL - SST/ISV - Institut des sciences de la vie
UCL - SST/LIBST - Louvain Institute of Biomolecular Science and Technology
Keywords Conditional mutagenesis ; CreERT2 ; Hoxa5 ; Site-specific recombinase ; Tomato reporter line
Links
  1. Birling Marie-Christine, Gofflot Françoise, Warot Xavier, Site-Specific Recombinases for Manipulation of the Mouse Genome, Transgenesis Techniques (2009) ISBN:9781603270182 p.245-263, 10.1007/978-1-60327-019-9_16
  2. Bradley Allan, Anastassiadis Konstantinos, Ayadi Abdelkader, Battey James F., Bell Cindy, Birling Marie-Christine, Bottomley Joanna, Brown Steve D., Bürger Antje, Bult Carol J., Bushell Wendy, Collins Francis S., Desaintes Christian, Doe Brendan, Economides Aris, Eppig Janan T., Finnell Richard H., Fletcher Colin, Fray Martin, Frendewey David, Friedel Roland H., Grosveld Frank G., Hansen Jens, Hérault Yann, Hicks Geoffrey, Hörlein Andreas, Houghton Richard, Hrabé de Angelis Martin, Huylebroeck Danny, Iyer Vivek, de Jong Pieter J., Kadin James A., Kaloff Cornelia, Kennedy Karen, Koutsourakis Manousos, Kent Lloyd K. C., Marschall Susan, Mason Jeremy, McKerlie Colin, McLeod Michael P., von Melchner Harald, Moore Mark, Mujica Alejandro O., Nagy Andras, Nefedov Mikhail, Nutter Lauryl M., Pavlovic Guillaume, Peterson Jane L., Pollock Jonathan, Ramirez-Solis Ramiro, Rancourt Derrick E., Raspa Marcello, Remacle Jacques E., Ringwald Martin, Rosen Barry, Rosenthal Nadia, Rossant Janet, Ruiz Noppinger Patricia, Ryder Ed, Schick Joel Zupicich, Schnütgen Frank, Schofield Paul, Seisenberger Claudia, Selloum Mohammed, Simpson Elizabeth M., Skarnes William C., Smedley Damian, Stanford William L., Francis Stewart A., Stone Kevin, Swan Kate, Tadepally Hamsa, Teboul Lydia, Tocchini-Valentini Glauco P., Valenzuela David, West Anthony P., Yamamura Ken-ichi, Yoshinaga Yuko, Wurst Wolfgang, The mammalian gene function resource: the international knockout mouse consortium, 10.1007/s00335-012-9422-2
  3. Di Meglio T., Kratochwil C. F., Vilain N., Loche A., Vitobello A., Yonehara K., Hrycaj S. M., Roska B., Peters A. H. F. M., Eichmann A., Wellik D., Ducret S., Rijli F. M., Ezh2 Orchestrates Topographic Migration and Connectivity of Mouse Precerebellar Neurons, 10.1126/science.1229326
  4. Ganat Y. M., Silbereis J., Cave C., Ngu H., Anderson G. M., Ohkubo Y., Ment L. R., Vaccarino F. M., Early Postnatal Astroglial Cells Produce Multilineage Precursors and Neural Stem Cells In Vivo, 10.1523/jneurosci.2532-06.2006
  5. Gofflot F, Wendling O, Chartoire N, Birling MC, Warot X, Auwerx J (2011) Characterization and validation of Cre-driver mouse lines. Curr Protoc Mouse Biol 1:1–15. doi: 10.1002/9780470942390.mo100103
  6. Grindley Nigel D.F., Whiteson Katrine L., Rice Phoebe A., Mechanisms of Site-Specific Recombination, 10.1146/annurev.biochem.73.011303.073908
  7. Hayashi Shigemi, McMahon Andrew P., Efficient Recombination in Diverse Tissues by a Tamoxifen-Inducible Form of Cre: A Tool for Temporally Regulated Gene Activation/Inactivation in the Mouse, 10.1006/dbio.2002.0597
  8. Hutlet B, Theys N, Coste C, Ahn MT, Doshishti-Agolli K, Lizen B, Gofflot F (2014) Systematic expression analysis of Hox genes at adulthood reveals novel patterns in the central nervous system. Brain Struct Funct. doi: 10.1007/s00429-014-0965-8
  9. Indra A. K., Warot X., Brocard J., Bornert J.-M., Xiao J.-H., Chambon P., Metzger D., Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ERT and Cre-ERT2 recombinases, 10.1093/nar/27.22.4324
  10. K�hbandner Susanne, Brummer Sabine, Metzger Daniel, Chambon Pierre, Hofmann Franz, Feil Robert, Temporally controlled somatic mutagenesis in smooth muscle, 10.1002/1526-968x(200009)28:1<15::aid-gene20>3.0.co;2-c
  11. Kuo Chay T., Mirzadeh Zaman, Soriano-Navarro Mario, Rašin Mladen, Wang Denan, Shen Jie, Šestan Nenad, Garcia-Verdugo Jose, Alvarez-Buylla Arturo, Jan Lily Y., Jan Yuh-Nung, Postnatal Deletion of Numb/Numblike Reveals Repair and Remodeling Capacity in the Subventricular Neurogenic Niche, 10.1016/j.cell.2006.10.041
  12. Madisen Linda, Zwingman Theresa A, Sunkin Susan M, Oh Seung Wook, Zariwala Hatim A, Gu Hong, Ng Lydia L, Palmiter Richard D, Hawrylycz Michael J, Jones Allan R, Lein Ed S, Zeng Hongkui, A robust and high-throughput Cre reporting and characterization system for the whole mouse brain, 10.1038/nn.2467
  13. Metzger Daniel, Chambon Pierre, Site- and Time-Specific Gene Targeting in the Mouse, 10.1006/meth.2001.1159
  14. Metzger D, Chambon P (2011) Generation of spatio-temporally controlled targeted somatic mutations in the mouse. Curr Protoc Mouse Biol 1:55–70. doi: 10.1002/9780470942390.mo100128
  15. Nagy Andras, Cre recombinase: The universal reagent for genome tailoring, 10.1002/(sici)1526-968x(200002)26:2<99::aid-gene1>3.0.co;2-b
  16. Nakamura Eiichiro, Nguyen Minh-Thanh, Mackem Susan, Kinetics of tamoxifen-regulated Cre activity in mice using a cartilage-specific CreERT to assay temporal activity windows along the proximodistal limb skeleton, 10.1002/dvdy.20892
  17. Novak Anton, Guo Caiying, Yang Wenyi, Nagy Andras, Lobe Corrinne G., Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon cre-mediated excision, 10.1002/1526-968x(200011/12)28:3/4<147::aid-gene90>3.0.co;2-g
  18. Pitulescu Mara E, Schmidt Inga, Benedito Rui, Adams Ralf H, Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice, 10.1038/nprot.2010.113
  19. Santagati F., Temporal requirement of Hoxa2 in cranial neural crest skeletal morphogenesis, 10.1242/dev.02078
  20. Shelton K.A., Cline J.M., Cann J.A., 17-β Estradiol reduces atherosclerosis without exacerbating lupus in ovariectomized systemic lupus erythematosus-susceptible LDLr−/− mice, 10.1016/j.atherosclerosis.2012.12.031
  21. Shoda T, Kato M, Harada R, Fujisato T, Okuhira K, Demizu Y, Inoue H, Naito M, Kurihara M (2015) Synthesis and evaluation of tamoxifen derivatives with a long alkyl side chain as selective estrogen receptor down-regulators. Bioorg Med Chem. doi: 10.1016/j.bmc.2015.05.002
  22. Smedley Damian, Salimova Ekaterina, Rosenthal Nadia, Cre recombinase resources for conditional mouse mutagenesis, 10.1016/j.ymeth.2010.12.027
  23. Tabariès Sébastien, Lemieux Margot, Aubin Josée, Jeannotte Lucie, Comparative analysis ofHoxa5 allelic series, 10.1002/dvg.20292
Bibliographic reference Lizen, Benoît ; Claus, Mélissa ; Jeannotte, Lucie ; Rijli , F.M. ; Gofflot, Françoise. Perinatal induction of Cre recombination with tamoxifen.. In: Transgenic Research, Vol. 24, p. 1065-1077 (2015)
Permanent URL http://hdl.handle.net/2078.1/165423