SMP30 silencing in vivo - stem cell biology (Sep/24/2008 )
I am working on a transplantation protocol in kidney and I want to silence smp30 gene of the host animal.
Is there a way to do it? Perhaps shRNA or something else?
I didn t find any commercial product targeting smp30 in vivo
thank you in advance
Vivo-Morpholinos are a good technique for kidney. When splice-modifying Vivo-Morpholinos were injected into mice at 12.5 mg/kg once per day for four days and then sacrificed on day 5 for RT-PCR analysis, over 90% of the targeted transcripts in kidney had underwent splice modification. See:
Li YF, Morcos PA. Design and Synthesis of Dendritic Molecular Transporter that Achieves Efficient in Vivo Delivery of Morpholino Antisense Oligo. Bioconjug Chem. 2008 Jul;19(7):1464-70. Epub 2008 Jun 20.
Morpholinos without delivery moieties have been used successfully in explanted organotypic kidney cultures.
Nikopoulos G, Adams T, Adams D, Oxburgh L, Prudovsky I, Verdi J. The use of Endo-Porter to deliver morpholinos in kidney organ culture. Biotechniques. 2008 Apr;44(4):547-9.
The oligos have far less off-target RNA interactions than RNAi-based techniques such as siRNA or shRNA.
Mende M, Christophorou NA, Streit A. Specific and effective gene knock-down in early chick embryos using morpholinos but not pRFPRNAi vectors. Mech Dev. 2008 Sep 3. [Epub ahead of print]
Morpholinos can target translation, splicing or miRNA maturation. The uncharged Morpholino oligo backbone does not trigger innate immune responses. They are commercially available with about two week turnaround for a custom synthesis from ordering to delivery. Targeting assistance is included.
ok thx. i am gonna check it out.