TY - JOUR

T1 - In vivo dsRNA production system for silencing the phytoene desaturase gene of potato Solanum tuberosum

AU - Ivanov, Artemii

AU - Golubeva, Tatiana

N1 - This work was supported by the Russian Science Foundation (grant No. 22-76-00037).

PY - 2024/2/22

Y1 - 2024/2/22

N2 - In this work, for the first time, an in vivo system was developed for the production of exogenous double-stranded RNA (dsRNA), complementary to the potato phytoene desaturase (pds) gene. For this purpose, a recombinant plasmid was constructed based on the L4440 vector; the pds gene fragment was inserted between two late T7 promoters, which allows the production of dsRNA. The resulting plasmid was used to transform the E. coli strain HT115, which is deficient in RNase III. This strain prevents the hydrolysis of the dsRNA produced in it. The presence of the required dsRNA fragments was confirmed by electrophoretic analysis. This system can be used as a positive control in experiments on regulating the activity of potato genes using spray-induced gene silencing (SIGS).

AB - In this work, for the first time, an in vivo system was developed for the production of exogenous double-stranded RNA (dsRNA), complementary to the potato phytoene desaturase (pds) gene. For this purpose, a recombinant plasmid was constructed based on the L4440 vector; the pds gene fragment was inserted between two late T7 promoters, which allows the production of dsRNA. The resulting plasmid was used to transform the E. coli strain HT115, which is deficient in RNase III. This strain prevents the hydrolysis of the dsRNA produced in it. The presence of the required dsRNA fragments was confirmed by electrophoretic analysis. This system can be used as a positive control in experiments on regulating the activity of potato genes using spray-induced gene silencing (SIGS).

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85188238158&origin=inward&txGid=1067571b767f1b1622cac8d741546f62

UR - https://www.mendeley.com/catalogue/4ef172f5-72d5-3773-865b-e3cbffa0a789/

U2 - 10.1051/e3sconf/202449404045

DO - 10.1051/e3sconf/202449404045

M3 - Conference article

VL - 494

JO - E3S Web of Conferences

JF - E3S Web of Conferences

SN - 2555-0403

M1 - 04045

T2 - 2023 International Conference on Ensuring Sustainable Development: Ecology, Energy, Earth Science and Agriculture

Y2 - 21 December 2023 through 22 December 2023

ER -