Tetraploid Complementation Assay

Gertsenstein, Marina

doi:10.1016/b978-0-12-394445-0.00009-6

Cited by 1 publication

(1 citation statement)

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“…37 One advantage of tetraploid complementation over blastocyst injection for producing knockout mouse lines is the vast difference in the time required for obtaining homozygous embryos. While the standard approach takes 8-12 mo, tetraploid complementation allows the production of virtually pure ES cell-derived homozygous embryos directly from the genetically altered cells (consecutively mutated on both alleles, which takes around 2-3 mo).…”

Section: Functional Analysis Of Lncrnas In Embryonic Developmentmentioning

confidence: 99%

The long non-coding RNAFendrrlinks epigenetic control mechanisms to gene regulatory networks in mammalian embryogenesis

2013

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Epigenetic control mechanisms determine active and silenced regions of the genome. It is known that the Polycomb Repressive Complex 2 (PRC2) and the Trithorax group/Mixed lineage leukemia (TrxG/Mll) complex are able to set repressive and active histone marks, respectively. Long non-coding RNAs (lncRNAs) can interact with either of these complexes and guide them to regulatory elements, thereby modifying the expression levels of target genes. The lncRNA Fendrr is transiently expressed in lateral mesoderm of mid-gestational mouse embryos and was shown to interact with both PRC2 and TrxG/Mll complexes in vivo. Gene targeting revealed that loss of Fendrr results in impaired differentiation of tissues derived from lateral mesoderm, the heart and the body wall, ultimately leading to embryonic death. Molecular data suggests that Fendrr acts via dsDNA/RNA triplex formation at target regulatory elements, and directly increases PRC2 occupancy at these sites. This, in turn, modifies the ratio of repressive to active marks, adjusting the expression levels of Fendrr target genes in lateral mesoderm. We propose that Fendrr also mediates long-term epigenetic marks to define expression levels of its target genes within the descendants of lateral mesoderm cells. Here we discuss approaches for lncRNA gene knockouts in the mouse, and suggest a model how Fendrr and possibly other lncRNAs act during embryogenesis.

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Section: Functional Analysis Of Lncrnas In Embryonic Developmentmentioning

confidence: 99%