ES cells already targeted with a T2A-Cre vector immediately downstream of the parvalbumin translational STOP codon, were re-targeted with a "T2A-Flpo" vector and a FLP-expressing plasmid to facilitate recombination. Correctly targeted ES cells had (from 5' to 3') a three amino acid linker sequence, an frt3 site within Pvalb intron 3, a partial Pvalb exon 4 sequence up to (but not including) the endogenous stop codon, a T2A sequence in-frame with the Pvalb coding sequence, a FlpO recombinase gene (a codon-optimized FLPe modified for translation in mammalian cells and higher recombination efficiency), the Pvalb 3' UTR sequence from exon 4, an attB site, a PGK-hygromycin-SV40polyA cassette (with a frt5 site in the hygromycin gene), and an attP site. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were bred to PhiC31-expressing mice (C57BL/6J congenic background; see JAX Stock No. 007743) to delete the attB/attP-flanked PGK-hygromycin-polyA cassette. Heterozygous mice have an FlpO recombinase expression pattern in the brain similar to endogenous Pvalb expression. (J:197021, J:219930)
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(129S6/SvEvTac x C57BL/6NCrl)F1
Targeted
Insertion
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1
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20

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hm: 纯合子
ht: 杂合子
cn: 条件基因型
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ot: 其他:半合子、不确定...
(F): 雌性
(M): 雄性
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