Human Transcription Factor Sirna Library Screening Apoptosis Related Of Sirna

RNA interference (RNAi) is a sequence-specific posttranscriptional gene silencing process mediated by double-stranded RNA. RNA interference occurs in a wide variety of organisms. Biologists have paid attention to RNAi since researchers found dsRNA can trigger the degradation of homologous mRNAs. Recently RNAi has become a commonly used technology to study the function of gene. Because long double-stranded RNA causes non-target-related induction of type 1 interferon (IFN), the application of RNAi technology is restricted to the lower organisms, such as C. elegans, Drosophila. The barrier was overcome with the observation that the sequence-specific RNAi pathway can be triggered by small interfering RNA (siRNA) without activating the IFN response. RNAi technology has been used not only to study the function of individual gene, but also to perform the RNAi-based genome-wide functional analysis for mammalian cells, which promotes the construction of siRNA library and high-throughput screen.The process of programmed cell death, or apoptosis, is generally characterized by distinct morphological characteristics and energy-dependent biochemical mechanisms. Apoptosis is considered a vital component of various processes including normal cell turnover, proper development and functioning of the immune system, hormone-dependent atrophy, embryonic development and chemical-induced cell death. Inappropriate apoptosis (either too little or too much) is the cause in many human disease including neurodegenerative diseases, ischemic damage, autoimmune disorders and many types of cancer. The ability to modulate the life or death of a cell is recognized for its immense therapeutic potential. Therefore, research continues to focus on the elucidation and analysis of apoptosis machinery and signaling pathways.We used the human transcription factor siRNA library in a screening for siRNA that can affect the cell apoptosis in jurkat cells. The cells were stably transfected with the siRNA library and then treated with rhTRAIL, cells with anti-apoptosis capacity would survive and propagate more than others and eventually become enriched in the cell population. After 3 round of selection, we found the cells transfected with the human transcription factor siRNA library did have anti-apoptosisi capacity than the cells transfected with the negative control plasmid. Then the siRNA expressing cassettes were recovered from the cells selected. These siRNA expressing cassettes were verified for their anti-apoptosis ability individually. Some of the siRNA expressing cassettes did have anti-apoptosis ability in jurkat cell. In summary, we used the huaman transcription factor siRNA library for high-throughput phenotype-driven screening, and multiple siRNAs that have anti-apoptosis ability were identified. Facilitating the further identification of novel apoptosis related genes.