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Lysine Specific Demethylase 1 (LSD1) knockout mouse

Invented by Shaun Cowley
Invented at University Of Leicester
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brachyury, a direct target gene of LSD1, is derepressed in Lsd1β-geo/β-geo gene trap embryos. (A) Quantitative RT-PCR shows upregulation of brachyury mRNA in Lsd1Δ3/Δ3 ES cells, with (day 0) and without (day 3) LIF. Values are expressed relative to the level of transcript in Lsd1Lox/Δ3 cells with LIF. (B) The Western blot shows an increase in brachyury protein levels in Lsd1Δ3/Δ3 ES cells with (day 0) and without (day 3) LIF. The fold induction of brachyury protein was calculated relative to Sin3A. (C) A schematic of the brachyury gene shows relative position of primers used for quantitative PCR following chromatin immunoprecipitation (ChIP). A region −600 bp from the transcriptional start site is enriched in anti-LSD1 ChIPs from Lsd1Lox/Δ3 but not Lsd1Δ3/Δ3 cells, showing specific enrichment of LSD1 protein (left panel). Loss of LSD1 protein results in increased histone H3 K4 dimethylation at the −600-bp region (right panel). (D) Quantitative RT-PCR analysis of e6.5 embryos. Homozygous (Lsd1β-geo/β-geo) embryos have a 20-fold increase in brachyury mRNA compared with wild-type and heterozygous (Lsd1+/β-geo) embryos. RasGRP3 transcription (5-fold) and CDA transcription (10-fold) are similarly increased in Lsd1β-geo/β-geo embryos. Assays were performed in triplicate with individual embryos of each genotype. Values are expressed relative to the level of transcript in wild-type embryos; mean values (n = 3) ± SEM are plotted.
A gene trap insertion inactivates Lsd1 and reveals embryo-specific expression. (A) A mouse embryonic stem (ES) cell line containing a gene trap vector in the Lsd1 gene locus (clone X102) was used to generate Lsd1+/β-geo mice. The gene trap vector, which consists of a splice acceptor site linked to a β-geo selectable marker, was found to be inserted into the 3rd intron of the Lsd1 gene on chromosome 4. (B) Schematic representation of LSD1. The position of the gene trap insertion (STOP), downstream of exon 3, truncates the LSD1 open reading frame within the SWIRM domain prior to the amine oxidase domain, which is essential for the catalytic activity of LSD1. Numbers indicate the positions of specific amino acids of the SWIRM, Tower, and amine oxidase domains. (C) The table shows the numbers and genotypes of viable pups and embryos isolated. Numbers of empty decidua are also listed. (D) Wild-type and Lsd1+/β-geo embryos isolated at 10.5 days postcoitum (dpc) were stained with X-Gal to detect β-galactosidase reporter gene activity approximating to LSD1 expression patterns. Similar X-Gal staining experiments were performed using embryos isolated at 8.5 dpc (E), 8.0 dpc (F), 6.5 dpc (G), and 3.5 dpc (H to J). The genotype of individual embryos is indicated. (K to M) X-Gal staining of blastocyst outgrowth cultures shows wild-type, Lsd1+/β-geo, and Lsd1β-geo/β-geo cells that were isolated at 3.5 dpc and cultured on gelatinized plates for 6 days. On day 6, blastocyst outgrowth cultures were stained with X-Gal to determine β-galactosidase expression and then genotyped.
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Catalogue Number 158387
Antigen/Gene or Protein Targets Lysine-Specific Demethylase 1
Synonyms LSD1, KDM1A, AOF2, KDM1, KIAA0601, LSD1
Disease Keywords Cleft palate, psychomotor retardation, and distinctive facial features (CPRF)
Model Knock-Out
Relevance Lysine-specific demethylase 1 (LSD1) acts as a coactivator or a corepressor of transcription, depending on the context, of histone H3 through demethylation of both 'Lys-4' (H3K4me) and 'Lys-9' (H3K9me). It demethylates mono- and dimethylated histone H3 as part of a larger complex. It essential for embryonic development in the mouse beyond embryonic day 6.5.
Production Details Embryos at the desired developmental stage were produced from timed Lsd1+/β-geo × Lsd1+/β-geo matings
Conditional No
Research Area Developmental Biology
Growth/Phenotype Keywords gene-trap
Notes Using this mouse model it was found that LSD1 regulates the expression and appropriate timing of key developmental regulators, as part of the LSD1/CoREST/HDAC complex, during early embryonic development.

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References: 1 entry

Foster et al. 2010. Mol Cell Biol. 30(20):4851-63. PMID: 20713442.

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References: 1 entry

Foster et al. 2010. Mol Cell Biol. 30(20):4851-63. PMID: 20713442.

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