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Comparative Silencing Effect of Different siRNA Fragments on Potato Virus X Coat Protein in Transient Transfection Assays

Comparative Silencing Effect of Different siRNA Fragments on Potato Virus X Coat Protein in Transient Transfection Assays

Anwar Khan1, Fareeha Nosheen2,4, Bushra Tabassum2*, Olawale Samuel Adeyinka2, Khurram Shehzad3, Naila Shahid2, Arif Muhammad Khan4 and Idrees Ahmad Nasir2

1Department of Microbiology, BUITEMS, Quetta, Pakistan
2Centre of Excellence in Molecular Biology, University of the Punjab, Lahore 53700, Pakistan
3Sahara Medical College, Narowal, Pakistan
4Department of Biotechnology, University of Sargodha, Sargodha, Pakistan

*      Corresponding author: bushratabassum.cemb@pu.edu.pk

Fig. 1.

Restriction digestion to confirm the expression constructs. (A) Restriction digestion of pVXC construct to reveal positivity of the clone. CP-PVX full length gene appears at 892 bp while pCDNA vector appears at 5.4 kb, (B) Restriction digestion of pSHX construct to reveal positivity of the clone. shRNA appears at 107 bp while pCDNA vector appears at 5.4 kb.

Fig. 2.

Knockdown of CP-PVX mRNA by target specific siRNA in cultured mammalian cells 24 h post transfection. (A) CP-PVX mRNA knockdown caused by siRNA1 in HepG2 cells. (B) CP-PVX mRNA knockdown caused by siRNA2 in HepG2 cells and (C) CP-PVX mRNA knockdown caused by siRNA3 in HepG2 cells. One-Way ANOVA and post Dunnet’s test were applied. The results showed that there was significant difference among control and different concentrations in standard value. (P < 0.05; n = 3).

Fig. 3.

Knockdown of CP-PVX by shRNA in HepG2 cells line. Four different traetments comprising of varied concentration of shRNA and target DNA were used to reveal mRNA suppression of coat protein gene of PVX. All samples were run in triplicate and normalized with GAPDH.

Pakistan Journal of Zoology

December

Vol. 53, Iss. 6, Pages 2001-2521

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