Computational Estimation and Experimental Verification of Off-Target Silencing during Posttranscriptional Gene Silencing in Plants

DOI PDF 被引用文献2件
  • Ping Xu
    Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401
  • Yuanji Zhang
    Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401
  • Li Kang
    Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401
  • Marilyn J. Roossinck
    Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401
  • Kirankumar S. Mysore
    Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401

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<jats:title>Abstract</jats:title> <jats:p>Successful application of posttranscriptional gene silencing (PTGS) for gene function study in both plants and animals depends on high target specificity and silencing efficiency. By computational analysis with genome and/or transcriptome sequences of 25 plant species, we predicted that about 50% to 70% of gene transcripts in plants have potential off-targets when used for PTGS that could obscure experimental results. We have developed a publicly available Web-based computational tool called siRNA Scan to identify potential off-targets during PTGS. Some of the potential off-targets obtained from this tool were tested by measuring the amount of off-target transcripts using quantitative reverse transcription-PCR. Up to 50% of the predicted off-target genes tested in plants were actually silenced when tested experimentally. Our results suggest that a high risk of off-target gene silencing exists during PTGS in plants. Our siRNA Scan tool is useful to design better constructs for PTGS by minimizing off-target gene silencing in both plants and animals.</jats:p>

収録刊行物

  • Plant Physiology

    Plant Physiology 142 (2), 429-440, 2006-08-18

    Oxford University Press (OUP)

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