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In silico Designing of Effective and Specific dsRNAs and siRNAs for Post-transcriptional Silencing of Whitefly, Bemisia tabaci Genes with Minimized Off-target Effects

ARCHNA SUHAG, ANKIT KUMARI, ANJALI JAIWAL, DARSHNA CHAUDHARY, PAWAN K. JAIWAL AND RANJANA JAIWAL*
Department of Zoology, Maharshi Dayanand University, Rohtak-124 001 (Haryana), India
*(e-mail: ranjana.jaiwal@gmail.com; Mobile: 99963 07959)
(Received: July 1, 2022; Accepted: August 7, 2022)

ABSTRACT

Whitefly (Bemisia tabaci) is a phloem-sap sucking and the most invasive insect-pest which transmits
several pathogenic viruses to the important crops worldwide causing a damage of billions of dollars per
year. Novel, effective and eco-friendly strategies are urgently required for their management. RNA
interference (RNAi) is a next generation bio-pesticide technology with high species-specificity and
environmentally-friendly. The four whitefly vital genes, namely, Vitellogenin receptor, Ryanodine receptor,
Nicotinic acetylcholine receptor beta 1 subunit and Laccase 1 that are important for the survival and
reproduction of whitefly were selected to design effective and specific dsRNAs and siRNAs for their
knockdown via RNAi. BLAST homology search was performed to remove off-target effects on the coding
sequences of whitefly selected genes sequences retrieved from the NCBI database. Potent siRNAs were
designed using i-score designer. Mean free-energy (MFE) calculated using RNA fold server was used to
identify sequences having lowest MFE and higher stability. The accessibility of mRNA target sites by
siRNAs was predicted by Sirna algorithm of S fold web server. The siRNAs selected in this study were
highly specific and efficient to silence whitefly genes with no or negligible off target effects.
Key words : Whitefly, management, RNAi, dsRNAs/siRNAs, off-target effects, in silico tools