PMID- 28585867 OWN - NLM STAT- MEDLINE VI - 24 IP - 1 TI - Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice. PG - 906-917 LA - eng PT - Journal Article PL - England TA - Drug Deliv JT - Drug delivery JID - 9417471 IS - 1521-0464 (Electronic) LID - 10.1080/10717544.2017.1333171 [doi] FAU - Oyama, Natsuko AU - Oyama N AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. FAU - Fuchigami, Yuki AU - Fuchigami Y AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. FAU - Fumoto, Shintaro AU - Fumoto S AUID- ORCID: http://orcid.org/0000-0002-2218-4547 AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. FAU - Sato, Megumu AU - Sato M AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. FAU - Hagimori, Masayori AU - Hagimori M AUID- ORCID: http://orcid.org/0000-0002-6718-2422 AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. FAU - Shimizu, Kazunori AU - Shimizu K AD - b Graduate School of Engineering , Nagoya University , Nagoya , Japan. FAU - Kawakami, Shigeru AU - Kawakami S AUID- ORCID: http://orcid.org/0000-0001-8765-2463 AD - a Graduate School of Biomedical Sciences , Nagasaki University , Nagasaki , Japan. IS - 1071-7544 (Linking) RN - 9007-49-2 (DNA) RN - EC 1.13.12.- (Luciferases) SB - IM MH - Animals MH - DNA/*genetics MH - Luciferases MH - Mice MH - Plasmids MH - Transfection MH - Transgenes OTO - NOTNLM OT - Gene delivery OT - fibroblast OT - kidney OT - naked pDNA OT - spatial distribution PMC - PMC8241128 DCOM- 20171212 LR - 20231107 DP - 2017 Nov AB - We have previously developed an efficient and safe transfection method for the kidney in mice: renal suction-mediated transfection. In this study, we verified the detailed characteristics of transgene expression and plasmid DNA (pDNA) in mice to develop therapeutic strategies and application to gene function analysis in the kidney. After naked pDNA was administered intravenously, the right kidney was immediately suctioned by a tissue suction device. We examined the spatial distribution of transgene expression and pDNA in the suctioned kidney using tissue clearing by CUBIC, ClearT2, and Scale SQ reagents. Spatial distribution analysis showed that pDNA was transfected into extravascular cells and sufficiently delivered to the deep renal cortex. In addition, we revealed that transgene expression occurred mainly in peritubular fibroblasts of the suctioned kidney by tissue clearing and immunohistochemistry. Next, we confirmed the periods of pDNA uptake and activation of transcription factors nuclear factor-κB and activator protein 1 by luciferase assays. Moreover, the use of a pCpG-free plasmid enabled sustained transgene expression in the suctioned kidney. In conclusion, analyses of the spatial distribution and immunostaining of the section suggest that pDNA and transgene expression occurs mainly in peritubular fibroblasts of the suctioned kidney. In addition, we clarified some factors for efficient and/or sustained transgene expression in the suctioned kidney.