Simple RNAi Vectors for Stable and Transient Suppression of Gene Function in Rice

To help identify the functions of genes in rice, we developed a Gateway vector, pANDA, for RNA interference of rice genes

Daisuke Miki; Ko Shimamoto

2004

Scholarcy highlights

  • RNA interference has been extensively used in various species to suppress gene function and is becoming a common tool for the functional analysis of the genome
  • Post-transcriptional gene silencing was first discovered in plants, and it is believed that its molecular mechanism is similar to that of RNAi observed in Caenorhabditis elegans, Drosophila, and mammals
  • The destination vector pANDA that we developed contains the maize ubiquitin promoter with an intron, which has been shown to give high expression of foreign genes in transgenic rice, and a 920 bp fragment of the coding region of the E. coli gus gene, which constituted a linker between two inverted repeats of the gene sequence derived from target genes
  • RNAi vectors for rice transformed with the phytoene desaturase gene-RNAi construct showed a clear albino phenotype, and the PDS mRNA levels were highly reduced. short-interfering RNA, a molecular marker for dsRNA-based gene silencing corresponding to the PDS sequence, was detected, confirming that the PDSRNAi construct made using the pANDA vector efficiently suppressed PDS function in transgenic rice plants
  • Since two size classes of siRNA have been reported in plants showing RNA silencing, we examined PDS siRNA after purifying low-molecular-weight RNA
  • When the GFP RNAi construct was cobombarded with the 35S-GFP, strong silencing of the green fluorescence was observed, while no difference in the expression of the control DsRed gene was observed. These results suggested that the RNAi vector based on the pANDA-mini was useful for transient suppression of gene function in rice leaf cells
  • These results suggested that the RNA interference vector based on the pANDA-mini was useful for transient suppression of gene function in rice leaf cells

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