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2021•IOP Conference Series Earth and Environmental ScienceOpen access

Silencing of Nicotiana benthamiana phytoendesaturase using dsRNA synthesized in vivo

В. А. Черенко, Е. А. Филипенко, Tatiana S. Golubeva

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Abstract

Abstract RNA interference (RNAi) using exogenous double-stranded RNA (dsRNA) has been used to silence the model gene of Nicotiana benthamiana phytoendesaturase. Here we report on an efficient technique for dsRNA synthesis using E. coli HT115 strain. This strain is deficient in RNase III, an enzyme that normally destroys most dsRNA in a bacterial cell and has been engineered to produce big quantities of dsRNA. We also used root treatment for dsRNA delivery to N. benthamiana plants. We found this method to be one of the most efficient ways to deliver dsRNA for plants.

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Abstract RNA interference (RNAi) using exogenous double-stranded RNA (dsRNA) has been used to silence the model gene of Nicotiana benthamiana phytoendesaturase. Here we report on an efficient technique for dsRNA synthesis using E. coli HT115 strain. This strain is deficient in RNase III, an enzyme that normally destroys most dsRNA in a bacterial cell and has been engineered to produce big quantities of dsRNA. We also used root treatment for dsRNA delivery to N. benthamiana plants. We found this method to be one of the most efficient ways to deliver dsRNA for plants.

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Available abstract

Abstract RNA interference (RNAi) using exogenous double-stranded RNA (dsRNA) has been used to silence the model gene of Nicotiana benthamiana phytoendesaturase. Here we report on an efficient technique for dsRNA synthesis using E. coli HT115 strain. This strain is deficient in RNase III, an enzyme that normally destroys most dsRNA in a bacterial cell and has been engineered to produce big quantities of dsRNA. We also used root treatment for dsRNA delivery to N. benthamiana plants. We found this method to be one of the most efficient ways to deliver dsRNA for plants.

Key concepts: Nicotiana benthamiana, RNA silencing, RNA interference, Gene silencing, RNA, RNase P, Biology, Ribonuclease III

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