Enhancing RNAi efficiency by inserting Nuclear factor-κB binding sequence into SiRNA expression cassette

RNAi based applications have been widely used to manipulate cellular phenotypes, analyze gene function and as a promising molecular therapy, however, there are rare reports in how to target the SiRNA expression constructs from the cytoplasm to nucleus. In this study, we inserted NF-κB binding sequence into the SiRNA expression cassette to enhance RNAi efficiency, tested this propose by use of tRNAval and human H1 promoters, and eGFP and luciferase as reporter protein. The results showed that inserting NF-κB binding sequence at the 3’-terminal of the SiRNA expression construct could enhance RNAi efficiency

Enhancing RNAi efficiency by inserting Nuclear factor-κB binding sequence into SiRNA expression cassette

RNAi based applications have been widely used to manipulate cellular phenotypes, analyze gene function and as a promising molecular therapy, however, there are rare reports in how to target the SiRNA expression constructs from the cytoplasm to nucleus. In this study, we inserted NF-κB binding sequence into the SiRNA expression cassette to enhance RNAi efficiency, tested this propose by use of tRNAval and human H1 promoters, and eGFP and luciferase as reporter protein. The results showed that inserting NF-κB binding sequence at the 3’-terminal of the SiRNA expression construct could enhance RNAi efficiency

Detecting Newcastle disease virus in combination of RT-PCR with red blood cell absorption

Reverse transcription-polymerase chain reaction (RT-PCR) has limited sensitivity when treating complicated samples, such as feces, waste-water in farms, and nucleic acids, protein rich tissue samples, all the factors may interfere with the sensitivity of PCR test or generate false results. In this study, we developed a sensitive RT-PCR, combination of red blood cell adsorption, for detecting Newcastle disease virus (NDV). One pair of primers which was highly homologous to three NDV pathotypes was designed according to the consensus nucleocapsid protein (NP) gene sequence. To eliminate the interfere of microbes and toxic substances, we concentrated and purified NDV from varied samples utilizing the ability of NDV binding red blood cells (RBCs). The RT-PCR coupled with red blood cell adsorption was much more sensitive in comparison with regular RT-PCR. The approach could also be used to detect other viruses with the property of hemagglutination, such as influenza viruses