Caption: Figure 4: p47phox gene knockdown
reduced bupivacaine-induced ROS overexpression and cell injury.
Caption: Figure 6: The effects of inhibiting autophagy by Beclin 1 gene knockdown
on EMT in AA-treated HKC cells.
Our results showed that the gene knockdown
efficiency and stability of the gene silencing effects were increased when the pooled siRNA fragments were used.
In the present study, application of RNAi was investigated in cultured buffalo granulosa cells by both gene specific siRNA and reporter vector based small hairpin RNA (shRNA) gene knockdown. The study demonstrates that siRNA transfection is well optimized for gene specific knockdown and thus strongly suppressed the expression of CYP19 gene and estradiol production in buffalo granulosa cells.
For the initial validation of plasmid and siRNA transfection, culture containing serum was used while rest of the experiments on CYP19 gene knockdown was done in serum free culture.
Further studies may determine the longevity of gene knockdown
and whether the effect could be incorporated into progeny or enhanced by multiple applications.
Fluorescence images of intracellular ROS in WT or CD38 knockdown H9c2 cells were captured using a fluorescent microscope with CM- H2DCFD staining when the cells were subjected to H/R, and a robust elevation of the intracellular ROS was shown in wild type cells, but not CD38 gene knockdown
cells (magnification, x100 scale bars, 200 [micro]m; *** p < 0.001).
We also detect the expression of HIF-1[alpha] (Figure 4(b)) and miR-210 (Figure 4(d)) to confirm the efficiency of gene knockdown
at the same time.
The use of ePAM-R for siRNA delivery was demonstrated by gene knockdown after transfecting high mobility group box-1 (HMGB1, a cytokine-like molecule) siRNA into H2O2- or NMDA-treated primary cortical cultures.
The G2 "rigid" dendrimers showed higher gene knockdown than the "flexible" analogues while maintaining less off-target effects than Lipofectamine.
Some examples of specific topics include derivation and manipulation of murine embryonic stem cells, efficient gene knockdowns
in human embryonic stem cells using lentiviral-based RNAi, ex vivo megakaryocyte expansion and platelet production from human cord blood stem cells, isolation and manipulation of mammalian neural stem cells in vitro, regeneration of skin and cornea by tissue engineering, template DNA- strand co-segregation and asymmetric cell division in skeletal muscle stem cells, derivation of contractile smooth muscle cells from embryonic stem cells, and reprogramming of liver to pancreas.