Abstract
Completion of whole genome sequencing in many plant species including economically important crop species not only opens up new opportunities but also imposes challenges for plant science research community. Functional validation and utilization of these enormous DNA sequences necessitate new or improved tools with high accuracy and efficiency. Of various tools, small RNA-mediated gene silencing platform plays an important and unique role in functional verification of plant genes and trait improvements. Artificial trans-acting small interfering RNA (atasiRNA) has emerged as a potent and specific gene silencing platform which overcomes major limitations of other small RNA silencing approaches including double-stranded RNA, artificial microRNA (amiRNA), and microRNA-induced gene silencing. To best utilize atasiRNA platform, it is essential to be able to test candidate atasiRNAs efficiently through either in vivo or in vitro validation approach. Very recently, a breakthrough has been made in developing a new method for in vitro screen of amiRNA candidates, named "epitope-tagged protein-based amiRNA screens". Such a screen can be readily employed to validate atasiRNA candidates and thus accelerate the deployment of atasiRNA technology. Therefore, atasiRNA as an emerging tool shall accelerate both plant biology study and crop genetic improvements including trait stacking.
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