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Viral Transduction

Laboratory Head
Mehdi Totonchi
totonchi
Technical Staff
Maryam Hajikaram (M.Sc)

RNA viruses, one of the diverse group of viruses, can infect a broad spectrum of cells from prokaryotic to eukaryotic. Among them, the Retrovirdae family is more applicable in molecular biology studies. Retroviruses that replicate in the host cell by the enzyme reverse transcriptase, are one of the interesting viruses used in molecular biology. These viruses can incorporate into the host genome after the production of DNA from its RNA genome, by an integrase enzyme. The retroviral genome consists of 3 ORFs, including gag, pol and env genes. The core and structural proteins of the virus are encoded by the gag sequence; the enzymes required for its life cycle including RT, protease and integrase are encoded by the pol sequence; and coat proteins are encoded by the env gene. Regulatory sequences of the retrovirus genome consist of two long terminal repeats (LTRs) on both sides of the coding sequence. 5’ LTR, which acts as a promoter and transcription start site and 3’ LTR which is involved in posttranscriptional processing (i.e., polyadenylation). The packaging signal placed just after the 5’LTR is responsible for packaging all sequences as a retrovirus genome. Lentiviruses are other members of the Retroviridae family with complex a genome that could infect both dividing and non-dividing cells. By replacing viral genes with our gene of interest and placing a transgene juxtaposed to the packaging signal on one vector, and engineering viral coding genes on the other vectors, it is possible to produce recombinant viruses carrying the gene of interest that capable of transduction into any target cell.

We began our work by first establishing human iPS cells in 2008. Currently, we Have the capability to produce iPS cells from any cell type.
Other works currently ongoing in the Viral Transduction lab include:

1.    RNAi trials for gene silencing and functional analysis studies,
2.    Over-expression and ectopic expression of genes for functional analysis
3.    PPI studies
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Generation of human induced pluripotent stem cells using viral transduction method.

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