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Here, we seek to develop a recombinant adenovirus system to optimally silence mouse BMP9 and then characterize the important role of BMP9 in osteogenic differentiation of MSCs.
Wnt10b siRNA was a pool of 3 target-specific 20-25-nt siRNAs designed to silence mouse Wnt10b gene (Santa-Cruz Biotechnology).
To validate ADMR as a potential therapeutic target, in vivo studies were conducted using neutral nanoliposomes to systemically deliver human siRNA to ADMR to silence human cancer cells and mouse siRNA to ADMR to silence mouse tumor stromal cells.
Because the transfection efficiency was low, we first tested whether our two miRNAs (which were designed to silence mouse TDP-43) could knockdown rat TDP-43 by transfecting our constructs into rat-2 cells, which can be transfected with above 90% efficiency.
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27mer RNA oligonucleotides for silencing mouse cathepsin B were purchased from IDT Technologies.
To test the efficiency of the short hairpin (sh) RNA vector in silencing mouse (m) Nr1d1 expression (Figure S1), HEK293T cells were transiently transfected in 100-mm plates at a confluence of 30% with 2μg of pcDNA3.1-mNr1d1 pcDNA3.1-mNr1d1pression vector for mouse Nr1d1.
The siRNA designed for silencing mouse KAP3 (M-047278-00-0010) was purchased from Dharmacon Research.
Using shRNA, we silenced mouse podocalyxin surface protein expression by 6.3-fold in 4T1-luc mouse mammary tumor cells (BALB/c mouse-derived).
Previously, we showed that knock-down of FPPS expression with siRNA prevents Ang II-induced hypertrophy in cultured cardiomyocytes [ 4], and now we report the establishment of a FPPS gene silencing mouse model using lentiviral vectors expressing siRNA.
Furthermore, the weight of the tumors was also remarkably decreased (p < 0.001) in SMC1A-silenced mice.
Sik1 silenced mice showed significantly faster re-entrainment when compared to the controls in both cycles.
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