Specific interference shRNA-expressing plasmids inhibit Hantaan virus infection in vitro and in vivo

Liu, Yuan-yuan; Chen, Liang-jun; Zhong, Yan; Shen, Meng-xin; Ma, Nian; Liu, Bing-yu; Luo, Fan; Hou, Wei; Yang, Zhan-qiu; Xiong, Hai-rong

shRNA design and selection Two individual targeting sites were selected for the S (GenBank accession number AF187082) and M genes (GenBank accession number AF276987) of HTNV. The design of the shRNA sequences was performed according to methods referred to in the literature [19] and involved the use of the webbased Block-iT RNAi Designer program (http://rnaidesigner. thermofisher.com/rnaiexpress/). A BLAST search (http:// www.ncbi.nlm.nih.gov/BLAST) was performed to exclude the possible homologous sequences. The sequences and positions of the shRNAs are illustrated in Table 1 . The designed structure contained the following sequences: 5'-AA-sense strand siRNA, CTTGCTTC (loop region); and antisense strand siRNA, TATAGTGA (T7-Oligo promoter)-3'. The target gene is presented in Table 1 . A negative control was chosen to target positions 1023 to 1043 of Leishmania CRK1, the sequence of which was 5'-AATCGGGCAGTTGTTTGAGAT-3'.