Perinatal induction of Cre recombination with tamoxifen
| Autor: | Benoit Lizen, Melissa Claus, Lucie Jeannotte, Françoise Gofflot, Filippo M. Rijli |
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| Rok vydání: | 2015 |
| Předmět: |
Male
medicine.drug_class Transgene Mice Transgenic Context (language use) Biology Andrology Mice Pregnancy Genetics medicine Recombinase Animals Site-specific recombinase technology Homeodomain Proteins Recombination Genetic Integrases Estrogen Antagonists Phosphoproteins medicine.disease Mice Inbred C57BL Perinatal Care Tamoxifen Animals Newborn Receptors Estrogen Estrogen Immunology Mice Inbred CBA Gestation Female Animal Science and Zoology Agronomy and Crop Science Transcription Factors Biotechnology medicine.drug |
| Zdroj: | Transgenic Research. 24:1065-1077 |
| ISSN: | 1573-9368 0962-8819 |
| DOI: | 10.1007/s11248-015-9905-5 |
| Popis: | 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-CreER(T2) 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. |
| Databáze: | OpenAIRE |
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