The use of Endo-Porter to deliver morpholinos in kidney organ culture.
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.
Nikopoulos
George N
GN
Maine Medical Center Research Institute, Center for Molecular Medicine, Scarborough, Maine 04074, USA.
Adams
Tamara L
TL
Adams
Derek
D
Oxburgh
Leif
L
Prudovsky
Igor
I
Verdi
Joseph M
JM
D000818:Animals; D016503:Drug Delivery Systems; D019793:Fluorescein / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D007668:Kidney / Q000166:cytology*; D051379:Mice; D008813:Mice, Inbred ICR; D008954:Models, Biological; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D009924:Organ Culture Techniques; D010455:Peptides; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors
C511217:Endo-Porter; D005456:Fluorescent Dyes; C486541:Maged1 protein, mouse; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D010455:Peptides; D012333:RNA, Messenger; D019793:Fluorescein