SARS-CoV (the SARS-Associated Coronavirus) was reported as a novel virus member in the coronavirus family, which was the cause of severe acute respiratory syndrome. Coronavirus replication occurs through a unique mechanism employing Leader sequence in the transcripts when initiating transcription from the genome. Therefore, we cloned the Leader sequence from SARS-CoV(BJ01), which is identical to that identified from SARS-CoV(HKU-39849), and constructed specific siRNA targeting the Leader sequence. Using EGFP and RFP reporter genes fused with the cloned SARS-CoV Leader sequence, we demonstrated that the siRNA targeting the Leader sequence decreased the mRNA abundance and protein expression levels of the reporter genes in 293T cells. By stably expressing the siRNA in Vero E6 cells, we provided data that the siRNA could effectively and specifically decrease the mRNA abundance of SARSCoV genes as analyzed by RT-PCR and Northern blot. Our data indicated that the siRNA targeting the Leader sequence inhibited the replication of SARS-CoV in Vero E6 cells by silencing gene expression. We further demonstrated, via transient transfection experiments, that the siRNA targeting the Leader sequence had a much stronger inhibitory effect on SARS-CoV replication than the siRNAs targeting the Spike gene or the antisense oligodeoxynucleotides did. This report provides evidence that targeting Leader sequence using siRNA could be a powerful tool in inhibiting SARS-CoV replication. Gene Therapy (2005) 12, 751-761.

Amplitude models are constructed to describe the resonance structure of and decays using pp collision data collected at centre-of-mass energies of 7 and 8 TeV with the LHCb experiment, corresponding to an integrated luminosity of 3.0 . The largest contributions to both decay amplitudes are found to come from axial resonances, with decay modes and being prominent in and , respectively. Precise measurements of the lineshape parameters and couplings of the , and resonances are made, and a quasi model-independent study of the resonance is performed. The coherence factor of the decays is calculated from the amplitude models to be , which is consistent with direct measurements. These models will be useful in future measurements of the unitary-triangle angle and studies of charm mixing and violation.