A more cost effective and rapid high percentage germ‐line transmitting chimeric mouse generation procedure via microinjection of 2‐cell, 4‐cell, and 8‐cell embryos with ES and iPS cells

Kraus, Petra; Leong, Geraldine; Tan, Valerie; Xing, Xing; Goh, Jie Wei; Yap, Sook Peng; Lufkin, Thomas

doi:10.1002/dvg.20627

Cited by 27 publications

(18 citation statements)

References 7 publications

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“…Pluripotent ESC can be derived from the ICM of blastocysts [8], [9]. Upon injection of pluripotent ESC into 2C–8C stage pre-implantation embryos [10] or blastocysts [11] chimeric organisms can be generated. The degree of chimerism is determined by the degree of pluripotency of any given injected ESC and its ability to outcompete cells of the host organism to achieve contribution to the chimeric organism and from an evolutionary point of view, most importantly its germ line [10].…”

Section: Introductionmentioning

confidence: 99%

Pleiotropic Functions for Transcription Factor Zscan10

Kraus

Sivakamasundari

et al. 2014

PLoS ONE

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The transcription factor Zscan10 had been attributed a role as a pluripotency factor in embryonic stem cells based on its interaction with Oct4 and Sox2 in in vitro assays. Here we suggest a potential role of Zscan10 in controlling progenitor cell populations in vivo. Mice homozygous for a Zscan10 mutation exhibit reduced weight, mild hypoplasia in the spleen, heart and long bones and phenocopy an eye malformation previously described for Sox2 hypomorphs. Phenotypic abnormalities are supported by the nature of Zscan10 expression in midgestation embryos and adults suggesting a role for Zscan10 in either maintaining progenitor cell subpopulation or impacting on fate choice decisions thereof.

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Section: Introductionmentioning

confidence: 99%

Pleiotropic Functions for Transcription Factor Zscan10

Kraus

Sivakamasundari

et al. 2014

PLoS ONE

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“…We achieved a targeting efficiency of 2% (3 of 148 clones) validated by Southern blotting (data not shown). The ES cells were injected into mouse embryos as previously described (Kraus et al, 2010) to generate germ line transmitting chimeras. The resulting heterozygous mice Dlx1as 4xPA(neo)/ þ were normal, viable and fertile and bred to the ZP3-Cre deleter strain (Lewandoski et al, 1997) for the removal of the loxP flanked selection cassette to generate Dlx1as 4xPA/ þ mice.…”

Section: Resultsmentioning

confidence: 99%

Making sense of Dlx1 antisense RNA

Kraus

Sivakamasundari

Lim

et al. 2013

Developmental Biology

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Long non-coding RNAs (lncRNAs) have been recently recognized as a major class of regulators in mammalian systems. LncRNAs function by diverse and heterogeneous mechanisms in gene regulation, and are key contributors to development, neurological disorders, and cancer. This emerging importance of lncRNAs, along with recent reports of a functional lncRNA encoded by the mouse Dlx5-Dlx6 locus, led us to interrogate the biological significance of another distal-less antisense lncRNA, the previously uncharacterized Dlx1 antisense (Dlx1as) transcript. We have functionally ablated this antisense RNA via a highly customized gene targeting approach in vivo. Mice devoid of Dlx1as RNA are viable and fertile, and display a mild skeletal and neurological phenotype reminiscent of a Dlx1 gain-of function phenotype, suggesting a role for this non-coding antisense RNA in modulating Dlx1 transcript levels and stability. The reciprocal relationship between Dlx1as and Dlx1 places this sense-antisense pair into a growing class of mammalian lncRNA-mRNA pairs characterized by inverse regulation.

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“…1). The Sox9 ?/floxFRTNeo ES cell clones were microinjected into 2-8 cell stage mouse embryos isolated from C57BL/6 J mice (Kraus et al 2010). Sox9 ?/floxFRTNeo chimeras were crossed to wild type C57BL6/6 J or CD1 mice to generate heterozygous mice.…”

Section: Generation Of the Sox9 Conditional Knockout Allelementioning

confidence: 99%

Generation of mice with a novel conditional null allele of the Sox9 gene

et al. 2011

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Sox9 is expressed in multiple tissues during mouse development and adulthood. Mutations in the Sox9 gene or changes in expression levels can be attributed to many congenital diseases. Heterozygous loss-of-function mutations in the human SOX9 gene cause Campomelic dysplasia, a semi-lethal skeletal malformation syndrome. Disruption of Sox9 by conventional gene targeting leads to perinatal lethality in heterozygous mice, hence hampering the feasibility to obtain the homozygous Sox9 null mice for in vivo functional studies. In this study, we generated a conditional allele of Sox9 (Sox9 ( tm4.Tlu )) by flanking exon 1 with loxP sites. Homozygous mice for the Sox9 ( tm4.Tlu ) allele (Sox9 ( flox/flox )) are viable, fertile and indistinguishable from wildtype (WT) mice, indicating that the Sox9 ( tm4.Tlu ) allele is a fully functional Sox9 allele. Furthermore, we demonstrated that Cre-mediated recombination using a Col2a1-Cre line resulted in specific ablation of Sox9 activity in cartilage tissues.

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A more cost effective and rapid high percentage germ‐line transmitting chimeric mouse generation procedure via microinjection of 2‐cell, 4‐cell, and 8‐cell embryos with ES and iPS cells

Cited by 27 publications

References 7 publications

Pleiotropic Functions for Transcription Factor Zscan10

Pleiotropic Functions for Transcription Factor Zscan10

Making sense of Dlx1 antisense RNA

Generation of mice with a novel conditional null allele of the Sox9 gene

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