RNAi Inhibition of Rabbit Hemorrhagic Disease Virus (RHDV) Gene Expression in CHO Cell Line


Article info

2007-09-29
2008-04-08
2008-04-08
145 - 165

Keywords

Abstract

Rabbit hemorrhagic disease virus (RHDV) causes great losses and is life threatening to both wild and domestic rabbits. This work represents the first step toward production of transgenic rabbits resistant to RHDV by RNAi through the inhibition of expression of a part of the RHDV genome and the consequent inhibition of its multiplication. This part of RHDV genome is available under the accession EU003582.1 in NCBI from the nucleotide 372 to 392 (McIntosh et al., 2007). Chinese Hamster Ovary (CHO) cell line was transfected with a plasmid harboring the RHDV target in the DNA segment coding for luciferase (fusion protein) and co-transfected with another plasmid harboring a DNA sequence designed to code for a homologous double stranded RNA sequence of 21 pb. This sequence acts as siRNA to inhibit the expression of the viral genomic part through RNAi. The series of co-transfection resulted in an inhibition level ranging from 82 to 87% of expression of the fusion protein and consequently of the RHDV target. These results are encouraging. However, further investigations are necessary to obtain more inhibition ex-vivo with validation in vivo using transgenic rabbits.

RNAi Inhibition of Rabbit Hemorrhagic Disease Virus (RHDV) Gene Expression in CHO Cell Line


معلومات المقال

2007-09-29
2008-04-08
2008-04-08
145 - 165

الكلمات الإفتتاحية

الملخص

Rabbit hemorrhagic disease virus (RHDV) causes great losses and is life threatening to both wild and domestic rabbits. This work represents the first step toward production of transgenic rabbits resistant to RHDV by RNAi through the inhibition of expression of a part of the RHDV genome and the consequent inhibition of its multiplication. This part of RHDV genome is available under the accession EU003582.1 in NCBI from the nucleotide 372 to 392 (McIntosh et al., 2007). Chinese Hamster Ovary (CHO) cell line was transfected with a plasmid harboring the RHDV target in the DNA segment coding for luciferase (fusion protein) and co-transfected with another plasmid harboring a DNA sequence designed to code for a homologous double stranded RNA sequence of 21 pb. This sequence acts as siRNA to inhibit the expression of the viral genomic part through RNAi. The series of co-transfection resulted in an inhibition level ranging from 82 to 87% of expression of the fusion protein and consequently of the RHDV target. These results are encouraging. However, further investigations are necessary to obtain more inhibition ex-vivo with validation in vivo using transgenic rabbits.

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