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MYMIV-AC2, a Geminiviral RNAi Suppressor Protein, Has Potential to Increase the Transgene Expression

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Abstract

Gene silencing is one of the limiting factors for transgene expression in plants. But the plant viruses have learnt to suppress gene silencing by encoding the protein(s), called RNA silencing suppressor(s) (RSS). Hence, these proteins could be used to overcome the limitation for transgene expression. The RNAi suppressors, namely HC-Pro and P19, have been shown to enhance the transgene expression but other RSS proteins have not been screened for similar role. Moreover, none of RSSs from the DNA viruses are known for enhancing the expression of transgenes. The Mungbean Yellow Mosaic India Virus (MYMIV) belonging to the genus Begomovirus within the family of Geminiviridae encodes an RSS called the AC2 protein. Here, we used AC2 to elevate the expression of the transgenes. Upon introduction of MYMIV-AC2 in the silenced GFP transgenic tobacco lines, by either genetic hybridisation or transgenesis, the GFP expression was enhanced several fold in F1 and T0 lines. The GFP-siRNA levels were much reduced in F1 and T0 lines compared with those of the initial parental silenced lines. The enhanced GFP expression was also observed at the cellular level. This approach was also successful in enhancing the expression of another transgene, namely topoisomeraseII.

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Abbreviations

GM:

Genetically modified

PTGS:

Post-translational gene silencing

TGS:

Transcriptional gene silencing

RNAi:

RNA interference

RdRP:

RNA-dependent RNA polymerase

MYMIV:

Mungbean Yellow Mosaic India Virus

VIGS:

Virus-induced gene silencing

GFP:

Green florescent protein

RSS:

RNA silencing suppressors

CaMV:

Cauliflower mosaic virus

MiRs:

Micro-RNAs

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Acknowledgements

We acknowledge Professor David C. Baulcombe, UK for providing the mGFP gene. We are also thankful to Professor M. V. Rajam, Delhi University South Campus for providing the TOPOII transgenic T2 tobacco lines. We thank Dr. M. N. Islam for his initial efforts to develop GFP-silenced lines. This research was supported by grants from International Centre for Genetic Engineering and Biotechnology (ICGEB) and Department of Biotechnology, New Delhi, India. The financial assistance of DBT and CSIR (India) to SM and SK are duly acknowledged.

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  1. Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi, 110067, India

    Jamilur Rahman, Sumona Karjee & Sunil Kumar Mukherjee

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  1. Jamilur Rahman
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  2. Sumona Karjee
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  3. Sunil Kumar Mukherjee
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Correspondence to Sunil Kumar Mukherjee.

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Rahman, J., Karjee, S. & Mukherjee, S.K. MYMIV-AC2, a Geminiviral RNAi Suppressor Protein, Has Potential to Increase the Transgene Expression. Appl Biochem Biotechnol 167, 758–775 (2012). https://doi.org/10.1007/s12010-012-9702-z

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