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Post-transcriptional gene silencing: Basic concepts and applications

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Abstract

Post-transcriptional gene silencing (PTGS)-mediated gene silencing exploits the cellular mechanism wherein transcripts having sequence similarity to the double-stranded RNA (dsRNA) molecules present in the cell will be subjected to degradation. PTGS is closely related to natural processes such as RNA-mediated virus resistance and cross-protection in plants. Gene silencing and the cellular machinery for affecting this phenomenon might have evolved as a natural protective measure against viral infection in plants. In PTGS, small interfering RNA (siRNA) molecules of 21–23 nucleotides length act as homology guides for triggering the systemic degradation of transcripts homologous to the siRNA molecules. PTGS phenomenon, first discovered in transgenic petunia plants harbouring chalcone synthase gene and termed co-suppression, has been subsequently exploited to target specific gene transcripts for degradation leading to manifestation of desirable traits in crop plants. Targeted gene silencing has been achieved either through the introduction of DNA constructs encoding dsRNA or antisense RNA or by deploying co-suppression constructs producing siRNAs against the transcript of interest. Understanding the mechanism of gene silencing has led to the development of several alternative strategies for inducing gene silencing in a precise and controlled way. This has paved the way for using PTGS as one of the chief functional genomics tools in plants and has helped in unraveling the mechanism of many cellular processes and identifying the focal points in pathways, besides, opening new vistas in genetic engineering of plants for human benefits. PTGS has shown great potential in silencing the deleterious genes efficiently so that value-added plant products could be obtained. Thus, PTGS has ushered in a new era in the genetic manipulation of plants for both applied and basic studies. In this review, we have outlined the basics of RNAi-mediated gene silencing and summarized the work carried out at our institute using this approach, as case studies. In particular, adopting RNAi-mediated gene silencing (a) as a method to restore fertility in transgenic male sterile lines developed based on orfH522 gene from sunflower PET1-CMS source, (b) as a tool to suppress the production of toxic proteins, ricin and RCA, in castor, and (c) as an approach to induce bud necrosis virus resistance in sunflower has been discussed. Examples from other plant systems also have been mentioned to exemplify the concept and utility of gene silencing in crop plants.

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Acknowledgements

The financial help by Indian Council of Agricultural Research, New Delhi, through a research grant (Code No. 030558005), Department of Biotechnology, New Delhi, through research grants (Code No. BT/PR9589/AGR/02/441/07 and BT/PR/9836/AGR/36/17/2007) to VDK for carrying out the research work on developing cell ablation system using orfH522 work; research Grant (Code SR/SO/BB-58/2005) by Department of Science Technology, New Delhi, to VDK to carry out research work on reducing ricin and RCA in castor through PTGS approaches; and the financial assistance from the Department of Biotechnology, New Delhi, through a research Grant (BT/PR9572/ARG/02/432/2007) to MS for work on developing transgenic sunflower lines are gratefully acknowledged. Research fellowship provided by CSIR to MAA is also acknowledged. The authors are also grateful to the Director, ICAR-IIOR, Hyderabad, for providing all the logistics support to carry out the research work presented in this article.

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  1. These authors have contributed equally to this work.

Authors and Affiliations

  1. ICAR-Indian Institute of Oilseeds Research (IIOR), Rajendranagar, Hyderabad, 500 030, India

    Mohd Ashraf Ashfaq, V Dinesh Kumar, P Soma Sekhar Reddy, Ch Anil Kumar, K Sai Kumar, N Narasimha Rao, M Tarakeswari & M Sujatha

  2. Department of Botany, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110 062, India

    Mohd Ashraf Ashfaq

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  1. Mohd Ashraf Ashfaq
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  2. V Dinesh Kumar
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  3. P Soma Sekhar Reddy
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  4. Ch Anil Kumar
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Correspondence to V Dinesh Kumar.

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This article is part of the Topical Collection: Genetic Intervention in Plants: Mechanisms and Benefits.

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Ashfaq, M.A., Dinesh Kumar, V., Soma Sekhar Reddy, P. et al. Post-transcriptional gene silencing: Basic concepts and applications. J Biosci 45, 128 (2020). https://doi.org/10.1007/s12038-020-00098-3

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