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The CRISPR/Cas9 system has been emerged to be a revolutionary genome engineering tool in various organisms recently.In most cases, the sgRNA target sites choose for gene modification were designed as GN19NGG or GGN 18NGG because of the limitation of promoters which make it difficult to apply the system into the research of short coding genes or small RNAs, such as microRNAs and piRNAs etc.Here we find the U6-2 promoter could drive the sgRNAs target start with all four nucleotide bases ATGC in model lepidopteran insect, Bombyx mori which means the further expansion of CRISPR site space.