Enhanced expression of human cDNA by phosphoglycerate kinase promoter-puromycin cassette in the mouse transthyretin locus

Li, Zhenghua; Zhao, Gang; Shen, Jingling; Araki, Kimi; Haruna, Kyoko; Inoue, Seiya; Wang, Jun; Yamamura, Ken Ichi

doi:10.1007/s11248-010-9389-2

Cited by 7 publications

(4 citation statements)

References 23 publications

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“…Northern Blot, Semiquantitative RT-PCR and Quantitative RT-PCR Analyses Total RNA was isolated from the brain, whole eyes, heart, lung, small intestine, testis, skeletal muscle, liver, and kidney. Northern blot and semiquantitative reverse transcriptionpolymerase chain reaction (RT-PCR) analyses were performed according to the methods described by Zhao et al 20 and Li et al, 23 respectively. The nucleotide sequence identity between the mouse and human Rbp4-coding regions is 87%.…”

Section: Generation Of Rbp4orf Knock-in Micementioning

confidence: 99%

Rescue of retinal morphology and function in a humanized mouse at the mouse retinol-binding protein locus

Liu

Suzuki

Shen

et al. 2017

Laboratory Investigation

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Retinol-binding protein RBP4 is the specific carrier for retinol in the blood. We previously produced a Rbp4-deficient (Rbp4) mouse that showed electroretinogram (ERG) abnormalities, accompanied by histological and electron-microscopic changes such as fewer synapses in the inner plexiform layer in the central retina. To address whether human RBP4 gene expression can rescue the phenotypes observed in Rbp4 mice, we produced a humanized (Rbp4) mouse with a human RBP4 open reading frame in the mouse Rbp4 locus using a Cre-mutant lox recombination system. In Rbp4 mice, the tissue-specific expression pattern of hRBP4orf was roughly the same as that of mouse Rbp4. ERG and morphological abnormalities observed in Rbp4 mice were rescued in Rbp4 mice as early as 7 weeks of age. The temporal expression pattern of hRBP4orf in the liver of Rbp4 mice was similar to that of mouse Rbp4 in Rbp4mice. In contrast, hRBP4orf expression levels in eyes were significantly lower at 6 and 12 weeks of age compared with mouse Rbp4 but were restored to the control levels at 24 weeks. The serum hRBP4 levels in Rbp4 mice were approximately 30% of those in Rbp4 at all ages examined. In accordance with this finding, the plasma retinol levels remained low in Rbp4 mice. Retinol accumulation in the liver occurred in control and Rbp4 mice but was higher in Rbp4 mice at 30 weeks of age. Mouse transthyretin expression was not altered in Rbp4 or Rbp4 mice. Taken together, 30% of the serum RBP4 level was sufficient to correct the abnormal phenotypes observed in Rbp4 mice.

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Section: Generation Of Rbp4orf Knock-in Micementioning

confidence: 99%

Rescue of retinal morphology and function in a humanized mouse at the mouse retinol-binding protein locus

Liu

Suzuki

Shen

et al. 2017

Laboratory Investigation

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“…Northern Blot, Semiquantitative RT-PCR, and qRT-PCR Analyses Northern blot and semiquantitative reverse transcriptionpolymerase chain reaction (RT-PCR) analyses were carried out as described by Zhao et al 16 and Li et al, 19 respectively. The nucleotide sequence identity between the mouse and human Rbp4 coding regions is~87%.…”

Section: Southern Blot Analyses For Isolation Of Targeted Es Cell Clonesmentioning

confidence: 99%

Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Shen

Shi

Suzuki

et al. 2016

Laboratory Investigation

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Retinol-binding protein 4 (RBP4) is a specific carrier for retinol in the blood. In hepatocytes, newly synthesized RBP4 associates with retinol and transthyretin and is secreted into the blood. The ternary transthyretin-RBP4-retinol complex transports retinol in the circulation and delivers it to target tissues. Rbp4-deficient mice in a mixed genetic background (129xC57BL/6J) have decreased sensitivity to light in the b-wave amplitude on electroretinogram. Sensitivity progressively improves and approaches that of wild-type mice at 24 weeks of age. In the present study, we produced Rbp4-deficient mice in the C57BL/6 genetic background. These mice displayed more severe phenotypes. They had decreased a-and b-wave amplitudes on electroretinograms. In accordance with these abnormalities, we found structural changes in these mice, such as loss of the peripheral choroid and photoreceptor layer in the peripheral retinas. In the central retinas, the distance between the inner limiting membrane and the outer plexiform layer was much shorter with fewer ganglion cells and fewer synapses in the inner plexiform layer. Furthermore, ocular developmental defects of retinal depigmentation, optic disc abnormality, and persistent hyaloid artery were also observed. All these abnormalities had not recovered even at 40 weeks of age. Our Rbp4-deficient mice accumulated retinol in the liver but it was undetectable in the serum, indicating an inverse relation between serum and liver retinol levels. Our results suggest that RBP4 is critical for the mobilization of retinol from hepatic storage pools, and that such mobilization is necessary for ocular development and visual function.

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“…The Neo in pEGFP-N1 was driven by its own promoter (SV-40), whereas the Neo in pEGFP-IRES-Neo was driven by the constitutive promoter (human elongation factor, EF) of EGFP separated by an upstream internal ribosome entry site (IRES) sequence. It had been reported that IRES-dependent second gene expression is significantly lower than cap-dependent first gene expression 38 , 39 . The lower expression of Neo may result in less bystander effects and non-transgenic cells will be killed by G418 in shorter time.…”

Section: Discussionmentioning

confidence: 98%

Efficient Generation of Transgenic Buffalos (Bubalus bubalis) by Nuclear Transfer of Fetal Fibroblasts Expressing Enhanced Green Fluorescent Protein

Luo

et al. 2018

Sci Rep

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The possibility of producing transgenic cloned buffalos by nuclear transfer of fetal fibroblasts expressing enhanced green fluorescent protein (EGFP) was explored in this study. When buffalo fetal fibroblasts (BFFs) isolated from a male buffalo fetus were transfected with pEGFP-N1 (EGFP is driven by CMV and Neo is driven by SV-40) by means of electroporation, Lipofectamine-LTX and X-tremeGENE, the transfection efficiency of electroporation (35.5%) was higher than Lipofectamine-LTX (11.7%) and X-tremeGENE (25.4%, P < 0.05). When BFFs were transfected by means of electroporation, more embryos from BFFs transfected with pEGFP-IRES-Neo (EGFP and Neo are driven by promoter of human elongation factor) cleaved and developed to blastocysts (21.6%) compared to BFFs transfected with pEGFP-N1 (16.4%, P < 0.05). A total of 72 blastocysts were transferred into 36 recipients and six recipients became pregnant. In the end of gestation, the pregnant recipients delivered six healthy calves and one stillborn calf. These calves were confirmed to be derived from the transgenic cells by Southern blot and microsatellite analysis. These results indicate that electroporation is more efficient than lipofection in transfecting exogenous DNA into BFFs and transgenic buffalos can be produced effectively by nuclear transfer of BFFs transfected with pEGFP-IRES-Neo.

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Enhanced expression of human cDNA by phosphoglycerate kinase promoter-puromycin cassette in the mouse transthyretin locus

Cited by 7 publications

References 23 publications

Rescue of retinal morphology and function in a humanized mouse at the mouse retinol-binding protein locus

Rescue of retinal morphology and function in a humanized mouse at the mouse retinol-binding protein locus

Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Efficient Generation of Transgenic Buffalos (Bubalus bubalis) by Nuclear Transfer of Fetal Fibroblasts Expressing Enhanced Green Fluorescent Protein

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