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Loss of lysyl oxidase-like 3 causes cleft palate and spinal deformity in mice.

Zhang J, Yang R, Liu Z, Hou C, Zong W, Zhang A, Sun X, Gao J - Hum. Mol. Genet. (2015)

Bottom Line: Failure of any one of these processes can result in embryonic malformation.We found that the obvious decrease of collagen cross-links in palate and spine that was induced by the lack of LOXL3 resulted in cleft palate and spinal deformity.The Loxl3 gene may be a candidate disease gene resulting in cleft palate and spinal deformity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Developmental Biology, School of Life Science, Shandong University, 27 Shanda Nanlu, Jinan 250100, China.

No MeSH data available.


Related in: MedlinePlus

Targeted inactivation of the Loxl3 gene. (A) Targeting strategy. Exons are numbered and depicted as white boxes. In the targeting construct, a loxP site was inserted upstream of exon 2 (containing the ATG start codon), and the FRT-neo-FRT-loxP cassette was inserted downstream of exon 2. (B) Identification of the targeted ES clones by long PCR with the indicated primers. Flox/+, targeted ES clone carrying the floxed Loxl3 allele; +/+, wild-type ES clone. (C) Identification of mouse genotypes by PCR with the indicated primers. +/+, wild-type mice; +/−, Loxl3+/− mice; −/−, Loxl3−/− mice. (D) Western blot analysis for LOXL3 and β-actin of Loxl3+/+, Loxl3+/−and Loxl3−/− fetuses at E14.5.
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DDV333F1: Targeted inactivation of the Loxl3 gene. (A) Targeting strategy. Exons are numbered and depicted as white boxes. In the targeting construct, a loxP site was inserted upstream of exon 2 (containing the ATG start codon), and the FRT-neo-FRT-loxP cassette was inserted downstream of exon 2. (B) Identification of the targeted ES clones by long PCR with the indicated primers. Flox/+, targeted ES clone carrying the floxed Loxl3 allele; +/+, wild-type ES clone. (C) Identification of mouse genotypes by PCR with the indicated primers. +/+, wild-type mice; +/−, Loxl3+/− mice; −/−, Loxl3−/− mice. (D) Western blot analysis for LOXL3 and β-actin of Loxl3+/+, Loxl3+/−and Loxl3−/− fetuses at E14.5.

Mentions: The mouse Loxl3 gene was inactivated by a two-step method consisting of homologous recombination in ES cells followed by Cre-recombination in mice (Fig. 1A). Homologous recombinants were identified by long PCR analyses (Fig. 1B). Deletion of exon 2 and the FRT-neo-FRT cassette by Cre-recombination led to the inactivation of the Loxl3 gene. Loxl3−/− mice were then generated by intercrossing Loxl3+/− mice. (Fig. 1C). Western blotting confirmed the loss of LOXL3 protein expression in the whole fetus from embryonic day 14.5 (E14.5) (Fig. 1D).Figure 1.


Loss of lysyl oxidase-like 3 causes cleft palate and spinal deformity in mice.

Zhang J, Yang R, Liu Z, Hou C, Zong W, Zhang A, Sun X, Gao J - Hum. Mol. Genet. (2015)

Targeted inactivation of the Loxl3 gene. (A) Targeting strategy. Exons are numbered and depicted as white boxes. In the targeting construct, a loxP site was inserted upstream of exon 2 (containing the ATG start codon), and the FRT-neo-FRT-loxP cassette was inserted downstream of exon 2. (B) Identification of the targeted ES clones by long PCR with the indicated primers. Flox/+, targeted ES clone carrying the floxed Loxl3 allele; +/+, wild-type ES clone. (C) Identification of mouse genotypes by PCR with the indicated primers. +/+, wild-type mice; +/−, Loxl3+/− mice; −/−, Loxl3−/− mice. (D) Western blot analysis for LOXL3 and β-actin of Loxl3+/+, Loxl3+/−and Loxl3−/− fetuses at E14.5.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4599675&req=5

DDV333F1: Targeted inactivation of the Loxl3 gene. (A) Targeting strategy. Exons are numbered and depicted as white boxes. In the targeting construct, a loxP site was inserted upstream of exon 2 (containing the ATG start codon), and the FRT-neo-FRT-loxP cassette was inserted downstream of exon 2. (B) Identification of the targeted ES clones by long PCR with the indicated primers. Flox/+, targeted ES clone carrying the floxed Loxl3 allele; +/+, wild-type ES clone. (C) Identification of mouse genotypes by PCR with the indicated primers. +/+, wild-type mice; +/−, Loxl3+/− mice; −/−, Loxl3−/− mice. (D) Western blot analysis for LOXL3 and β-actin of Loxl3+/+, Loxl3+/−and Loxl3−/− fetuses at E14.5.
Mentions: The mouse Loxl3 gene was inactivated by a two-step method consisting of homologous recombination in ES cells followed by Cre-recombination in mice (Fig. 1A). Homologous recombinants were identified by long PCR analyses (Fig. 1B). Deletion of exon 2 and the FRT-neo-FRT cassette by Cre-recombination led to the inactivation of the Loxl3 gene. Loxl3−/− mice were then generated by intercrossing Loxl3+/− mice. (Fig. 1C). Western blotting confirmed the loss of LOXL3 protein expression in the whole fetus from embryonic day 14.5 (E14.5) (Fig. 1D).Figure 1.

Bottom Line: Failure of any one of these processes can result in embryonic malformation.We found that the obvious decrease of collagen cross-links in palate and spine that was induced by the lack of LOXL3 resulted in cleft palate and spinal deformity.The Loxl3 gene may be a candidate disease gene resulting in cleft palate and spinal deformity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Developmental Biology, School of Life Science, Shandong University, 27 Shanda Nanlu, Jinan 250100, China.

No MeSH data available.


Related in: MedlinePlus