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10.1371/journal.pone.0168638 http://scihub22266oqcxt.onion/10.1371/journal.pone.0168638 C5333796!5333796 !28253301     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=28253301 &cmd=llinks): Failed to open stream: HTTP request failed! 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Targeting gene expression to specific cells of kidney tubules in vivo, using adenoviral promoter fragments |pdf=|usr=}}{{28253301 }} gab.com Text Targeting gene expression to specific cells of kidney tubules in vivo, using adenoviral promoter fragments JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=28253301 #FOAvip Although techniques for cell-specific gene expression via viral transfer have advanced, many challenges (e.g., viral vector design, transduction of genes into specific target cells) still remain. We investigated a novel, simple methodology for using adenovirus transfer to target specific cells of the kidney tubules for the expression of exogenous proteins. We selected genes encoding sodium-dependent phosphate transporter type 2a (NPT2a) in the proximal tubule, sodium-potassium-2-chloride cotransporter (NKCC2) in the thick ascending limb of Henle (TALH), and aquaporin 2 (AQP2) in the collecting duct. The promoters of the three genes were linked to a GFP-coding fragment, the final constructs were then incorporated into an adenovirus vector, and this was then used to generate gene-manipulated viruses. After flushing circulating blood, viruses were directly injected into the renal arteries of rats and were allowed to site-specifically expression in tubule cells, and rats were then euthanized to obtain kidney tissues for immunohistochemistry. Double staining with adenovirus-derived EGFP and endogenous proteins were examined to verify orthotopic expression, i.e. "adenovirus driven NPT2a-EGFP and endogenous NHE3 protein", "adenovirus driven NKCC2-EGFP and endogenous NKCC2 protein" and "adenovirus driven AQP2-EGFP and endogenous AQP2 protein". Owing to a lack of finding good working anti-NPT2a antibody, an antibody against a different protein (sodium-hydrogen exchanger 3 or NHE3) that is also specifically expressed in the proximal tubule was used. Kidney structures were well-preserved, and other organ tissues did not show EGFP staining. Our gene transfer method is easier than using genetically engineered animals, and it confers the advantage of allowing the manipulation of gene transfer after birth. This is the first method to successfully target gene expression to specific cells in the kidney tubules. This study may serve as the first step for safe and effective gene therapy in the kidney tubule diseases. Twit Text FOAVip Targeting gene expression to specific cells of kidney tubules in vivo, using adenoviral promoter fragments ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=28253301 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28253301 #FOAvip English Wikipedia <ref>{{Cite pmid|28253301 }}</ref> Targeting gene expression to specific cells of kidney tubules in vivo, using adenoviral promoter fragments #MMPMID28253301 Watanabe S ; Ogasawara T ; Tamura Y ; Saito T ; Ikeda T ; Suzuki N ; Shimosawa T ; Shibata S ; Chung UI ; Nangaku M ; Uchida S PLoS One 2017[]; 12 (3 ): e0168638 PMID28253301 show ga Although techniques for cell-specific gene expression via viral transfer have advanced, many challenges (e.g., viral vector design, transduction of genes into specific target cells) still remain. We investigated a novel, simple methodology for using adenovirus transfer to target specific cells of the kidney tubules for the expression of exogenous proteins. We selected genes encoding sodium-dependent phosphate transporter type 2a (NPT2a) in the proximal tubule, sodium-potassium-2-chloride cotransporter (NKCC2) in the thick ascending limb of Henle (TALH), and aquaporin 2 (AQP2) in the collecting duct. The promoters of the three genes were linked to a GFP-coding fragment, the final constructs were then incorporated into an adenovirus vector, and this was then used to generate gene-manipulated viruses. After flushing circulating blood, viruses were directly injected into the renal arteries of rats and were allowed to site-specifically expression in tubule cells, and rats were then euthanized to obtain kidney tissues for immunohistochemistry. Double staining with adenovirus-derived EGFP and endogenous proteins were examined to verify orthotopic expression, i.e. "adenovirus driven NPT2a-EGFP and endogenous NHE3 protein", "adenovirus driven NKCC2-EGFP and endogenous NKCC2 protein" and "adenovirus driven AQP2-EGFP and endogenous AQP2 protein". Owing to a lack of finding good working anti-NPT2a antibody, an antibody against a different protein (sodium-hydrogen exchanger 3 or NHE3) that is also specifically expressed in the proximal tubule was used. Kidney structures were well-preserved, and other organ tissues did not show EGFP staining. Our gene transfer method is easier than using genetically engineered animals, and it confers the advantage of allowing the manipulation of gene transfer after birth. This is the first method to successfully target gene expression to specific cells in the kidney tubules. This study may serve as the first step for safe and effective gene therapy in the kidney tubule diseases. |Adenoviridae/*genetics [MESH] |Gene Expression [MESH] |Genetic Vectors/genetics [MESH] |HEK293 Cells [MESH] |Humans [MESH] |Kidney Tubules/*cytology/*metabolism [MESH] |Promoter Regions, Genetic/*genetics [MESH] |Safety [MESH]Targeting gene expression to specific cells of kidney tubules in vivo,(epmc).pdf DeepDyve Pubget Overpricing