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Histone deacetylase 2 (HDAC 2) knockout mouse embryonic stem cell line

Invented by Shaun Cowley
Invented at University Of Leicester
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Generation of HDAC1 and HDAC2 conditional knockout ES cell lines. (A) An E14 ES cell line constitutively expressing a Cre/Estrogen Receptor (CreER) fusion from the ROSA26 locus was used to generate homozygous conditional knockout alleles for HDAC1 and HDAC2. Both copies of exon 2 were flanked by LoxP sites, using consecutive rounds of gene targeting. (B) Southern blots showing wild-type and exon 2 targeted HDAC1Lox/Lox and HDAC2Lox/Lox loci. Addition of 4-hydroxy tamoxifen (OHT) activates the CreER fusion and induces deletion of exon2 (HDAC1Δ2/Δ2, HDAC2Δ2/Δ2) in ES cells; >95% recombination is observed after 24 h. (C) Quantitative Western blot shows ligand-inducible deletion of HDAC1 and -2 protein in whole-cell extracts from HDAC1Lox/Lox and HDAC2Lox/Lox ES cells, respectively. Cells were cultured for up to 10 days (0–3 days in the presence of OHT). α-Tubulin was used to normalize protein loading. Blots were visualized and quantitated using a LiCOR scanner. Data are representative of three independently tested clones for each genotype.
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Catalogue Number 158384
Antigen/Gene or Protein Targets Histone deacetylase 2
Parental Line E14 ES cell line expressing an inducible Cre/Estrogen Receptor (CreER) construct from the endogenous ROSA26 locus
Synonyms HDAC2
Tissue Embryo
Disease Keywords Embryo Lethality
Model Knock-Out
Relevance Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Histone deacetylases (HDAC) 1 and 2 are highly similar enzymes that help regulate chromatin structure as the core catalytic components of corepressor complexes. Although tissue-specific deletion of HDAC1 and HDAC2 has demonstrated functional redundancy, germ-line deletion of HDAC1 in the mouse causes early embryonic lethality, whereas HDAC2 does not.
Production Details E14 ES cells, expressing a CreER fusion protein from the ROSA26 locus, were used to generate HDAC1Lox/Lox; CreER and HDAC2Lox/Lox; CreER cell lines by consecutive rounds of gene targeting.
Conditional Yes
Conditional Description Induction of Cre activity by addition of 4-hydroxy tamoxifen (OHT) to the growth media resulted in complete recombination of each allele and deletion of exon 2 (HDAC1Δ2/Δ2 or HDAC2 Δ2/Δ2) within 24 h
Research Area Developmental Biology, Reproductive Biology
Notes Loss of exon 2 via the conditional KO disrupts the ORF of both HDAC1 and HDAC2 such that a premature stop codon is introduced in exon 3. Following this deletion of exon 2 a further 4–5 days of culture are required before HDAC1 and HDAC2 protein levels are reduced below 10% of those of control cells.
Conditional deletion of HDAC1 or HDAC2 does not inhibit the growth or differentiation capacity of ES cells. Loss of HDAC1, but not HDAC2, Causes Enhanced Differentiation of Embryoid Bodies.

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References: 1 entry

Dovey et al. 2010. Proc Natl Acad Sci U S A. 107(18):8242-7. PMID: 20404188.

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References: 1 entry

Dovey et al. 2010. Proc Natl Acad Sci U S A. 107(18):8242-7. PMID: 20404188.

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