Establishment of transgenic lettuce plants producing potentially anti-hypertensive shRNA

Gerasymenko I.M., Kleschevnikov V.V., Kedlian V.R., Sakhno L.O., Arbuzova I.A., Sheludko Y.V., Dosenko V.E., Kuchuk N.V.

SUMMARY. Development of RNAi-based therapeutics is a fast growing field of pharmaceutical industry. Using plants for production of pharmaceutically valuable siRNAs may have significant advantages of cost-effectiveness, scalability and low risk of contamination with human pathogens. If edible plant species are genetically engineered to synthesize siRNAs, the costly stage of target product purification may be omitted. We describe the establishment of transgenic lettuce plants producing shRNA targeting delta isoform of protein kinase C (PKC-delta), an effective target for RNAi-based treatment of arterial hypertension. Transgenic lettuce plants were obtained by Agrobacterium-mediated transformation with genetic constructs harboring antiPKC and scrambled (control) shRNA genes. The presence of transgenes was proved by PCR analysis, and the accumulation of antiPKC shRNA was estimated using RT-qPCR technique. Six transgenic lettuce lines showed varying levels of antiPKC shRNA expression with the highest value reaching 14 ± 9 % of highly abundant endogenous lettuce micro RNA (miR156a), or 12.7 fmol/g dry weight. Plants carrying either antiPKC or scrambled shRNA genes flowered normally, but did not produce seeds. The described transgenic lettuce plants accumulating antiPKC siRNA are the subject for animal testing and can be considered as a raw material for the development of novel antihypertensive drugs.

Key words: RNAi, shRNA, arterial hypertension, delta isoform of protein kinase C, transgenic plants, Lactuca sativa

Tsitologiya i Genetika 2017, vol. 51, no. 1, pp. 3-11

E-mail: i-gerasimenko ukr.net

Gerasymenko I.M., Kleschevnikov V.V., Kedlian V.R., Sakhno L.O., Arbuzova I.A., Sheludko Y.V., Dosenko V.E., Kuchuk N.V. Establishment of transgenic lettuce plants producing potentially anti-hypertensive shRNA, Tsitol Genet., 2017, vol. 51, no. 1, pp. 3-11.

In "Cytology and Genetics":
I. M. Gerasymenko, V. V. Kleschevnikov, V. R. Kedlian, L. O. Sakhno, I. A. Arbuzova, Y. V. Sheludko, V. E. Dosenko, N. V. Kuchuk Establishment of transgenic lettuce plants producing potentially antihypertensive ShRNA, Cytol Genet., 2017, vol. 51, no. 1, pp. 1–7
DOI: 10.3103/S0095452717010054