The Use of Endo-Porter to Deliver Morpholinos in Kidney Organ Culture
Author(s) -
George N. Nikopoulos,
Tamara L. Adams,
Derek Adams,
Leif Oxburgh,
Igor Prudovsky,
Joseph M. Verdi
Publication year - 2008
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112725
Subject(s) - morpholino , mesenchyme , kidney , biology , organ culture , microbiology and biotechnology , kidney development , computational biology , hek 293 cells , cell culture , gene , bioinformatics , genetics , gene knockdown , mesenchymal stem cell , in vitro , embryonic stem cell
Cellular interactions in development of the kidney are used as a model of reciprocal inductive events between epithelium and mesenchyme. Time- and labor-intensive methods have been developed to study this phenomenon. For example, in mice, the targeted disruption of genes in vivo has been used to modify the genetic program directing kidney development. However, gene targeting is a resource-intensive approach and alternative strategies for gene and protein modification in the kidney need to be developed. Herein, we have developed an efficient system for the delivery of antisense morpholino to alter normal protein expression. We describe the use of Endo-Porter to effectively deliver morpholinos to all parts and regions of the kidney explant. Also, we definitively show via confocal microscopy and Western blot analysis that the use of Endo-Porter in delivering antisense morpholinos is robust throughout the entire kidney explant, providing efficient suppression of protein expression. This method saves time and cost when compared with targeted disruption and is an improvement upon previous kidney organ culture methods.
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