Related Experiment Video
Updated: Jun 10, 2026
Evaluation of the Efficacy And Toxicity of RNAs Targeting HIV-1 Production for Use in Gene or Drug Therapy
Published on: September 5, 2016
[Detection of IFN Response of Non-Specific Effects on RNAi.]
Yonggang Liang1, Hongyu Liu, Baoxing Liu
1Department of Thoracic and Caridovascular Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
Background:
The RNA interference (RNAi) approach is an efficient and widely used method for silencing specific genes and exploring their functions. However, numerous studies have highlighted the non-specific effects of RNAi experiments and the IFN response is one of the most common non-specific effects. In this research, we will study how to detect the IFN response in RNAi experiments.
Methods:
Five different FLJ20420 siRNAs was transfected into human lung adenocarinoma cell line A549 using LipofectamineTM2000. The alternation of the interferon stimulated genes (ISGs) expression profile was analysed by real-time PCR and microarray assay.
Results:
(1) Different FLJ20420 siRNA had different effect on silencing FLJ20420 mRNA expression in A549 cells, and FLJsiRNA-1 and -4 had the better effects to knockdown the expression of FLJ20420 (80% and 90% down-regulation, respectively); (2) In analyzed 16 major ISGs, the expression of 14 of these genes was significantly increased in A549-FLJsiRNA-1 cells (P <0.05), while only 2 of these genes had a significant change in A549-FLJ-siRNA-4 cells. These results indicated an IFN response existed in A549-FLJ-siRNA-1 cells. (3) Microarray assay indicated that the expression of 51 ISGs were increased over 2 times in A549-FLJ-siRNA-1 cells, while only the expression of 6 ISGs were over 2-time in A549-FLJ-siRNA-4 cells. These results confirmed that there was an IFN response of non-specific effects of RNAi in A549 cells transfected with FLJ-siRNA-1.
Conclusions:
In RNAi experiments, siRNA can not only silence the target genes, but also may activate IFN response. The analysis of the expression change of the major ISGs by real-time PCR is a useful approach to explore the IFN response in RNAi experiments.
Related Concept Videos
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
01:20Types of RNA
RNA Performs Diverse...
01:23Types of RNA
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
02:15Experimental RNAi
02:30siRNA - Small Interfering RNAs
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...