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Flp deleter mice

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Flp-deleter mice are a type of genetically modified mouse that express the Flp recombinase enzyme. The Flp recombinase enzyme is capable of removing DNA sequences that are flanked by Flp recognition target (FRT) sites. This allows for the targeted deletion of specific DNA sequences in the mouse genome.

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Lab products found in correlation

C57bl 6j female, by Jackson ImmunoResearch (2 mentions) Albumin cre mice, by Jackson ImmunoResearch (1 mentions) D12492, by Research Diets (1 mentions) 107787 aav8, by Addgene (1 mentions) Cd4 cre mice, by Jackson ImmunoResearch (1 mentions)

5 protocols using flp deleter mice

1

Myeloid-specific Orai1 Knockout Mice

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Stim1fl/fl animals were purchased from Jackson Laboratory (stock No. 023350) and bred with Lyz2-cre animals (Jackson Laboratory, stock No. 004781) for two generations. Targeting of murine Orai1 was performed by flanking exon 2 with LoxP sites by homologous recombination in AB2.2 (129SvEv) embryonic stem (ES) cells. Exon 2 encodes for 201 a.a. out of a total of 304 a.a. of Orai1 protein. G418-resistant clones were screened by PCR for homologous recombination at both homology arms. Chimeric mice with floxed Orai1 alleles were generated by blastocyst injection of heterozygous Orai1fl/+ ES cell clones. Founder Orai1fl/+ mice were bred with Flp-deleter mice (Jackson Laboratory) to remove the neomycin resistance gene cassette. Orai1fl/fl mice were backcrossed to C57/BL6/J mice for at least 10 generations and then bred with Lyz2-cre mice to generate myeloid-specific deletion of Orai1. All mice were maintained in pathogen-free barrier facilities and used in accordance with protocols approved by the Institutional Animal Care and Use Committee at the UCLA.
Srikanth S., Woo J.S., Wu B., El-Sherbiny Y.M., Leung J., Chupradit K., Rice L., Seo G.J., Calmettes G., Ramakrishna C., Cantin E., An D.S., Sun R., Wu T.T., Jung J.U., Savic S, & Gwack Y. (2019). The Ca2+ sensor STIM1 regulates type I interferon response by retaining the signaling adaptor STING at the endoplasmic reticulum. Nature immunology, 20(2), 152-162.
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2

Generation of Pten S398A Mutant Mice

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Chimeric mice were produced by microinjection of independent PtenS398A ES cell clones into E3.5 C57BL/6J blastocysts and transferred to ICR pseudopregnant foster mothers. Chimeric males were mated with C57BL/6J females (Jackson Laboratory). Germ line transmission of the mutant allele was confirmed by PCR and Southern blot analysis of tail DNA from mice with an agouti coat color. The PGK-Neo cassette was removed by crossing with Flp-deleter mice (Jackson Laboratory stock #009086) [30 (link)] and PCR genotyping and sequence validation of recombination at the FRT sites.
All experimental procedures strictly adhered to the Canadian Council on Animal Care guidelines.
Bassi C., Fortin J., Snow B.E., Wakeham A., Ho J., Haight J., You-Ten A., Cianci E., Buckler L., Gorrini C., Stambolic V, & Mak T.W. (2021). The PTEN and ATM axis controls the G1/S cell cycle checkpoint and tumorigenesis in HER2-positive breast cancer. Cell Death and Differentiation, 28(11), 3036-3051.
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3

Generation of Irf2 Knockout Mice

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Chimeric mice were produced by microinjection of independent mIrf2+/− ES cell clones into E3.5 C57BL/6J blastocysts and transferred to pseudopregnant foster mothers. Chimeric males were mated with C57Bl/6J females (Jackson Laboratory). Germ line transmission of the mutant allele was confirmed by PCR analysis of tail DNA from mice with an agouti coat color. The PGK-Neo cassette was removed from mouse Irf2+/− mice by crossing with Flp-deleter mice (Jackson Laboratory stock #009086) (Farley et al., 2000 (link)) and PCR genotyping and sequence validation of recombination at the FRT sites.
Lukhele S., Abd Rabbo D., Guo M., Shen J., Elsaesser H.J., Quevedo R., Carew M., Gadalla R., Snell L.M., Mahesh L., Ciudad M.T., Snow B.E., You-Ten A., Haight J., Wakeham A., Ohashi P.S., Mak T.W., Cui W., McGaha T.L, & Brooks D.G. (2022). The transcription factor IRF2 drives interferon-mediated CD8+ T cell exhaustion to restrict anti-tumor immunity. Immunity, 55(12), 2369-2385.e10.
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4

Liver-Specific Slc25a47 Knockout Mouse Model

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All the animal experiments in this study were performed in compliance with protocols approved by the Institutional Animal Care and Use Committee at Beth Israel Deaconess Medical Center. The Slc25a47 floxed (Slc25a47flox/flox) mouse was generated by in vitro fertilization of homozygous sperm (UC David) from Slc25a47tm1a (EUCOMM)Hmgu targeting exons 5 and 6 of the Slc25a47 gene in C57BL/6J background. A floxed LacZ-neomycin cassette on the Tm1a allele was removed using a flippase (FLP)/Frt deletion by breeding Slc25a47flox/flox with FLP deleter mice (Jackson Laboratory, Stock No. 009086). Slc25a47flox/flox mice were bred with Albumin Cre mice (Jackson Laboratory, Stock No. 003574) to generate liver-specific Slc25a47 deletion mice (Slc25a47Alb-Cre). Mice were kept under a 12-h:12-h light–dark cycle at ambient temperature (22 to 23 °C) and had free access to food and water. Mice were maintained on a regular chow diet or fed with a high-fat diet (60% fat, D12492, Research Diets) starting from 6 wk of age for 6 wk. All mice were fasted for 6 h before killing. To acutely deplete Slc25a47, we injected 7-wk-old Slc25a47flox/flox mice with 1.5 × 1011 genome copies of AAV8-TBG-Cre (Addgene, 107787-AAV8) or AAV8-TBG-null (control, Addgene, 105536-AAV8) through tail vein injection.
Yook J.S., Taxin Z.H., Yuan B., Oikawa S., Auger C., Mutlu B., Puigserver P., Hui S, & Kajimura S. (2023). The SLC25A47 locus controls gluconeogenesis and energy expenditure. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2216810120.
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5

Targeted Deletion of CRACR2A Gene

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Targeting of CRACR2A gene was performed by flanking exons 3 and 4 with LoxP sites by homologous recombination in AB2.2 (129SvEv) embryonic stem (ES) cells. Aberrant splicing of exon 2 to exon 5 causes a shift in reading frame resulting in truncation of the protein. Deletion of exons 3 and 4 would result in loss of expression of both the isoforms of CRACR2A. G418 resistant clones were PCR screened for homologous recombination at both the homology arms. Chimeric mice with floxed CRACR2A alleles were generated by blastocyst injection of heterozygous CRACR2Afl/+ ES cell clones. Founder CRACR2Afl/+ chimeric mice were bred with Flp-deleter mice (Jackson Laboratories) to remove neomycin resistance gene cassette. CRACR2A mice were backcrossed to C57BL/6 mice for at least 10 generations and then bred with CD4Cre mice (Jackson Laboratories) to generate T cell-specific deletion of CRACR2A. All animals were maintained in pathogen-free barrier facilities and used in accordance with protocols approved by the Institutional Animal Care and Use Committee at the University of California, Los Angeles.
Srikanth S., Kim K.D., Gao Y., Woo J.S., Ghosh S., Calmettes G., Paz A., Abramson J., Jiang M, & Gwack Y. (2016). A large Rab GTPase encoded by CRACR2A is a component of novel subsynaptic vesicles that transmit T cell activation signals. Science signaling, 9(420), ra31.
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