Utilizing siRNAs and shRNAs to Modulate Cellular Signaling

Small interfering RNAs (siRNAs) constructs used were obtained as the siGENOME SMARTpool reagents (Dharmacon, Lafayette, CO). The siRNA constructs used were: CREB1 siRNAs were purchased from Genepharma (Shanghai, China). ERK1 siGENOME SMARTpool (M-003592-03-0005), ERK2 siGENOME SMARTpool (M-003555-04-0005), and non-targeting siRNA pool (D-001206-13-20) as control. Transfection of siRNAs was carried out as described previously [65 (link)]. The shRNA targeting Noxa were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). Generation of lentiviruses expressing Noxa or control small hairpin RNAs (shRNAs) were as described previously [30 (link)].

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Liu Y.L., Lai F., Wilmott J.S., Yan X.G., Liu X.Y., Luan Q., Guo S.T., Jiang C.C., Tseng H.Y., Scolyer R.A., Jin L, & Zhang X.D. (2014). Noxa upregulation by oncogenic activation of MEK/ERK through CREB promotes autophagy in human melanoma cells. Oncotarget, 5(22), 11237-11251.

Publication 2014

siGENOME SMARTpool reagents

Erk1 Erk2 Lentiviruses Shrnas Sirnas Transfection

Corresponding Organization :

Other organizations : University of Newcastle Australia, Royal Prince Alfred Hospital, University of Sydney, Shanxi Provincial Cancer Hospital

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Variable analysis

independent variables

SiRNA constructs targeting CREB1, ERK1, and ERK2
ShRNA targeting Noxa

dependent variables

Not explicitly mentioned

control variables

Non-targeting siRNA pool
Control small hairpin RNAs (shRNAs)

controls

Positive control: Non-targeting siRNA pool
Negative control: Control small hairpin RNAs (shRNAs)

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