Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice

Oyama, Natsuko; Fuchigami, Yuki; Fumoto, Shintaro; Sato, Megumu; Hagimori, Masayori; Shimizu, Kazunori; Kawakami, Shigeru

doi:10.1080/10717544.2017.1333171

Cited by 15 publications

(21 citation statements)

References 43 publications

(86 reference statements)

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“…To identify the transfected cell types, we performed immunohistochemical analysis after the transfection of ZsGreen1-N1. In the heart transfected by the suction method, transgene expression was observed in the CD31-positive endothelial cells but not in cardiomyocytes ( Fig 5B); however, when the suction method was applied to the kidney, transgene expression was observed in the pericytes but not in the endothelial cells of the kidney [10]. The penetration of macromolecules is prevented by a vascular wall composed of tight junctions, adhesion junctions, and basement membranes [18,19].…”

Section: Discussionmentioning

confidence: 96%

“…In a translational research, the kinetics of a transfected protein should be analyzed. We and other group have previously reported that pDNA https://doi.org/10.1371/journal.pone.0228203.t003 removed CG motif (CpG free pDNA) could be used for sustained gene expression [10,[29][30].…”

Section: Discussionmentioning

confidence: 99%

“…Total mRNA was extracted from the minced organ by using RNeasy Fibrous Tissue Mini Kit (74704; Qiagen, Hilden, Germany). Thereafter, reverse transcription of mRNA and realtime PCR using SYBR Premix Ex Taq (RR039A; Takara Bio) was performed as described previously [10]. The sequences of primers used in the present study are shown in Table 2.…”

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

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Tissue suction-mediated gene transfer to the beating heart in mice

et al. 2020

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We previously developed an in vivo site-specific transfection method using a suction device in mice; namely, a tissue suction-mediated transfection method (tissue suction method). The aim of this study was to apply the tissue suction method for cardiac gene transfer. Naked plasmid DNA (pDNA) was intravenously injected in mice, followed by direct suction on the beating heart by using a suction device made of polydimethylsiloxane. We first examined the effects of suction conditions on transgene expression and toxicity. Subsequently, we analyzed transgene-expressing cells and the transfected region of the heart. We found that heart suction induced transgene expression, and that −75 kPa and −90 kPa of suction achieved high transgene expression. In addition, the inner diameter of the suction device was correlated with transgene expression, but the pressure hold time did not change transgene expression. Although the tissue suction method at −75 kPa induced a transient increase in the serum cardiac toxicity markers at 6 h after transfection, these markers returned to normal at 24 h. The cardiac damage was also analyzed through the measurement of hypertrophic gene expression, but no significant differences were found. In addition, the cardiac function monitored by echocardiography remained normal at 11 days after transfection. Immunohistochemical analysis revealed that CD31-positive endothelial cells coexpressed the ZsGreen1-N1 reporter gene. In conclusion, the tissue suction method can achieve an efficient and safe gene transfer to the beating heart in mice.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

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Tissue suction-mediated gene transfer to the beating heart in mice

et al. 2020

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“…Previously, we developed a tissue suction-mediated transfection method (suction method) in various mouse tissues using a single hole device with following hole dimensions: inner diameter 3 mm, outer diameter 5 mm and height 3 mm. [13][14][15][16] This tissue suction device can be loaded onto the head of an endoscope, making the tissue suction method potentially applicable in the clinic, using a novel medical device. Because we previously reported that transgene expression levels were dependent on the number of suctions, 13) transgene expression would be expected to increase in proportion to the number of holes in the suction device.…”

Section: Discussionmentioning

confidence: 99%

“…The types of physical stimuli tested include hydrodynamics, [2][3][4][5][6][7] mechanical massage, 8,9) pressure [10][11][12] and suction. [13][14][15][16] We described a liver suction-mediated transfection method (liver suction method), using a suction device with a single hole (single hole device) fabricated from polydimethylsiloxane. We believe that suction pressure waveform can be controlled by using a pressure-controlled computer system and the tissue suction device can be uploaded onto the head of an endoscope.…”

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confidence: 99%

Determining Transgene Expression Characteristics Using a Suction Device with Multiple Hole Adjusting a Left Lateral Lobe of the Mouse Liver

Haraguchi

Fuchigami

Kawaguchi

et al. 2018

Biological & Pharmaceutical Bulletin

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We developed a tissue suction-mediated transfection method (suction method) as a relatively reliable and less invasive technique for in vivo transfection. In this study, we determined hepatic transgene expression characteristics in the mouse liver, using a suction device, collecting information relevant to gene therapy and gene functional analysis by the liver suction method. To achieve high transgene expression levels, we developed a suction device with four holes (multiple hole device) and applied it to the larger portion of the left lateral lobe of the mouse liver. Hepatic transfection with physical stimuli was potentially controlled by activator protein-1 (AP-1) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). We examined the spatial distribution of transgene expression in the suctioned lobe by 2-dimensional imaging with histochemical staining and 3-dimensional multicolor deep imaging with tissue clearing methods. Through monitoring spatial distribution of transgene expression, the liver suction method was used to efficiently transfect extravascular hepatocytes in the suction-deformable upper lobe of the liver. Moreover, long-term transgene expression, at least 14 d, was achieved with the liver suction method when cytosine-phosphate-guanine (CpG)-free plasmid DNA was applied.

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Polyprenol-Based Lipofecting Agents for In Vivo Delivery of Therapeutic DNA to Treat Hypertensive Rats

et al. 2020

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Development of efficient vectors for transfection is one of the major challenges in genetic engineering. Previous research demonstrated that cationic derivatives of polyisoprenoids (PTAI) may serve as carriers of nucleic acids. In the present study, the effectiveness of two PTAI-based formulations (PTAI-6-8 and 10-14) was investigated and compared to the commercial reagents. The purpose of applied gene therapy was to enhance the expression of vascular endothelial growth factor (VEGF-A) in the renal medulla of spontaneously hypertensive rats (SHR) and to test its potential as a novel antihypertensive intervention. In the first part of the study (in vitro), we confirmed that PTAI-based lipoplexes efficiently transfect XC rat sarcoma cells and are stable in 37 °C for 7 days. In the in vivo experiments, we administered selected lipoplexes directly to the kidneys of conscious SHR (via osmotic pumps). There were no blood pressure changes and VEGF-A level in renal medulla was significantly higher only for PTAI-10-14-based formulation. In conclusion, despite the promising results, we were not able to achieve VEGF-A expression level high enough to verify VEGF-A gene therapy usefulness in SHR. However, results of our study give important indications for the future development of PTAI-based DNA carriers and kidney-targeted gene delivery.

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Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice

Cited by 15 publications

References 43 publications

Tissue suction-mediated gene transfer to the beating heart in mice

Tissue suction-mediated gene transfer to the beating heart in mice

Determining Transgene Expression Characteristics Using a Suction Device with Multiple Hole Adjusting a Left Lateral Lobe of the Mouse Liver

Polyprenol-Based Lipofecting Agents for In Vivo Delivery of Therapeutic DNA to Treat Hypertensive Rats

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