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  1. Article ; Online: Preparation, Assembly, and Transduction of Transgenic Elements Using Mosaic Analysis with Dual Recombinases (MADR).

    Rincon Fernandez Pacheco, David / Sabet, Sara / Breunig, Joshua J

    STAR protocols

    2020  Volume 1, Issue 3, Page(s) 100199

    Abstract: This protocol focuses on the cloning and stable integration of sequences of interest by the use of a mosaic analysis with dual recombinases (MADR) plasmid that includes fusion proteins or independent proteins under the control of 2A peptide or IRES ... ...

    Abstract This protocol focuses on the cloning and stable integration of sequences of interest by the use of a mosaic analysis with dual recombinases (MADR) plasmid that includes fusion proteins or independent proteins under the control of 2A peptide or IRES elements. Additionally, we describe how to generate a neural stem cell culture from Gt(ROSA)26Sort
    MeSH term(s) Animals ; Animals, Genetically Modified ; Electroporation ; Genetic Engineering/methods ; Genetic Vectors/chemistry ; Genetic Vectors/genetics ; Green Fluorescent Proteins/genetics ; Luminescent Proteins/genetics ; Mice ; Plasmids/genetics ; Recombinases/genetics ; Recombinases/metabolism ; Transfection ; Red Fluorescent Protein
    Chemical Substances Luminescent Proteins ; Recombinases ; Green Fluorescent Proteins (147336-22-9)
    Language English
    Publishing date 2020-12-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2020.100199
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Single-blind clinical trial: Sperm selection based on capacity to pass through cumulus oophorous column improves ICSI outcomes.

    Sabet, Sara / Najafi, Mohammad Hasan / Tavalaee, Marziyeh / Sadeghi, Niloofar / Nasr-Esfahani, Mohammad H

    Andrology

    2021  Volume 9, Issue 5, Page(s) 1560–1570

    Abstract: Background: Sperm selection procedures for future strategies that aim to select normal spermatozoa with intact DNA to improve intracytoplasmic sperm injection (ICSI) outcomes are in early developing stage.: Objectives: The objective is to find out ... ...

    Abstract Background: Sperm selection procedures for future strategies that aim to select normal spermatozoa with intact DNA to improve intracytoplasmic sperm injection (ICSI) outcomes are in early developing stage.
    Objectives: The objective is to find out whether the sperm selection procedure based on the ability of spermatozoa to traverse the cumulus cells could improve clinical outcomes of ICSI technique in infertile couples with male factor etiology.
    Materials and methods: For this single-blind clinical trial, mature metaphase II oocytes were retrieved from 150 couples with male factor infertility, male age lower than 45 years and female age under 38 years. These couples were divided into two groups. In control group (n = 75), spermatozoa processed by density gradient centrifugation (DGC) were used to inject the oocytes. In the study group (n = 75), the oocytes were divided into sibling groups. In one sibling group (DGC), the oocytes were inseminated with DGC-processed spermatozoa while in the other group (DGC-CC), they were inseminated with DGC-processed spermatozoa that passed cumulus oophorous column.
    Results: Mean fertilization and embryo quality were significantly higher in DGC-CC group compared to DGC and control group. In addition, mean of chemical pregnancy (52.27% vs. 34.14%; p = 0.05), clinical pregnancy based on sac (52.27% vs. 32.92%; p = 0.03), clinical pregnancy with heart beat (52.27% vs. 25.60%; p = 0.003) and ongoing pregnancy (43.18% vs. 21.95%; p = 0.02) rates were significantly higher in DGC-CC group compared to control group.
    Conclusion: Sperm selection based on integrated systems such as DGC and ability to pass through cumulus oophorous column could improve clinical outcomes of ICSI in couples with male factor infertility.
    MeSH term(s) Adult ; Centrifugation, Density Gradient ; Cumulus Cells/physiology ; Female ; Fertilization in Vitro/methods ; Humans ; Infertility, Male/physiopathology ; Infertility, Male/therapy ; Male ; Middle Aged ; Oocytes ; Pregnancy ; Pregnancy Rate ; Single-Blind Method ; Sperm Injections, Intracytoplasmic/methods ; Spermatozoa/physiology ; Treatment Outcome
    Language English
    Publishing date 2021-06-10
    Publishing country England
    Document type Journal Article ; Randomized Controlled Trial ; Research Support, Non-U.S. Gov't
    ZDB-ID 2696108-8
    ISSN 2047-2927 ; 2047-2919
    ISSN (online) 2047-2927
    ISSN 2047-2919
    DOI 10.1111/andr.13043
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 6983: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Rapid Generation of Somatic Mouse Mosaics with Locus-Specific, Stably Integrated Transgenic Elements.

    Kim, Gi Bum / Rincon Fernandez Pacheco, David / Saxon, David / Yang, Amy / Sabet, Sara / Dutra-Clarke, Marina / Levy, Rachelle / Watkins, Ashley / Park, Hannah / Abbasi Akhtar, Aslam / Linesch, Paul W / Kobritz, Naomi / Chandra, Swasty S / Grausam, Katie / Ayala-Sarmiento, Alberto / Molina, Jessica / Sedivakova, Kristyna / Hoang, Kendy / Tsyporin, Jeremiah /
    Gareau, Daniel S / Filbin, Mariella G / Bannykh, Serguei / Santiskulvong, Chintda / Wang, Yizhou / Tang, Jie / Suva, Mario L / Chen, Bin / Danielpour, Moise / Breunig, Joshua J

    Cell

    2020  Volume 179, Issue 1, Page(s) 251–267.e24

    Abstract: In situ transgenesis methods such as viruses and electroporation can rapidly create somatic transgenic mice but lack control over copy number, zygosity, and locus specificity. Here we establish mosaic analysis by dual recombinase-mediated cassette ... ...

    Abstract In situ transgenesis methods such as viruses and electroporation can rapidly create somatic transgenic mice but lack control over copy number, zygosity, and locus specificity. Here we establish mosaic analysis by dual recombinase-mediated cassette exchange (MADR), which permits stable labeling of mutant cells expressing transgenic elements from precisely defined chromosomal loci. We provide a toolkit of MADR elements for combination labeling, inducible and reversible transgene manipulation, VCre recombinase expression, and transgenesis of human cells. Further, we demonstrate the versatility of MADR by creating glioma models with mixed reporter-identified zygosity or with "personalized" driver mutations from pediatric glioma. MADR is extensible to thousands of existing mouse lines, providing a flexible platform to democratize the generation of somatic mosaic mice. VIDEO ABSTRACT.
    MeSH term(s) Animals ; Brain Neoplasms/genetics ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Gene Targeting/methods ; Genetic Loci/genetics ; Glioma/genetics ; HEK293 Cells ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Mutagenesis, Insertional/methods ; Neural Stem Cells/metabolism ; Recombinases/metabolism ; Transfection ; Transgenes/genetics
    Chemical Substances Recombinases
    Language English
    Publishing date 2020-08-07
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 187009-9
    ISSN 1097-4172 ; 0092-8674
    ISSN (online) 1097-4172
    ISSN 0092-8674
    DOI 10.1016/j.cell.2019.08.013
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1167: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)
    Zs.MG 58: Show issues

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  4. Article: Rapid Generation of Somatic Mouse Mosaics with Locus-Specific, Stably Integrated Transgenic Elements

    Kim, Gi Bum / Rincon Fernandez Pacheco, David / Saxon, David / Yang, Amy / Sabet, Sara / Dutra-Clarke, Marina / Levy, Rachelle / Watkins, Ashley / Park, Hannah / Abbasi Akhtar, Aslam / Linesch, Paul W / Kobritz, Naomi / Chandra, Swasty S / Grausam, Katie / Ayala-Sarmiento, Alberto / Molina, Jessica / Sedivakova, Kristyna / Hoang, Kendy / Tsyporin, Jeremiah /
    Gareau, Daniel S / Filbin, Mariella G / Bannykh, Serguei / Santiskulvong, Chintda / Wang, Yizhou / Tang, Jie / Suva, Mario L / Chen, Bin / Danielpour, Moise / Breunig, Joshua J

    Cell. 2019 Sept. 19, v. 179, no. 1

    2019  

    Abstract: In situ transgenesis methods such as viruses and electroporation can rapidly create somatic transgenic mice but lack control over copy number, zygosity, and locus specificity. Here we establish mosaic analysis by dual recombinase-mediated cassette ... ...

    Abstract In situ transgenesis methods such as viruses and electroporation can rapidly create somatic transgenic mice but lack control over copy number, zygosity, and locus specificity. Here we establish mosaic analysis by dual recombinase-mediated cassette exchange (MADR), which permits stable labeling of mutant cells expressing transgenic elements from precisely defined chromosomal loci. We provide a toolkit of MADR elements for combination labeling, inducible and reversible transgene manipulation, VCre recombinase expression, and transgenesis of human cells. Further, we demonstrate the versatility of MADR by creating glioma models with mixed reporter-identified zygosity or with “personalized” driver mutations from pediatric glioma. MADR is extensible to thousands of existing mouse lines, providing a flexible platform to democratize the generation of somatic mosaic mice.[Display omitted]
    Keywords electroporation ; enzymes ; glioma ; human cell lines ; loci ; mice ; mutants ; mutation ; transgenes ; transgenesis ; transgenic animals ; viruses
    Language English
    Dates of publication 2019-0919
    Size p. 251-267.e24.
    Publishing place Elsevier Inc.
    Document type Article
    ZDB-ID 187009-9
    ISSN 1097-4172 ; 0092-8674
    ISSN (online) 1097-4172
    ISSN 0092-8674
    DOI 10.1016/j.cell.2019.08.013
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Bonn / Germany

    Z 75/702: Show issues
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