Gene Analysis

Masters
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Science
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Retroviral based gene delivery system is widely used in gene therapy but there are many evidences indicating its catastrophic role in tumor induction, as it can integrate into different places of genome and cause insertion mutagenesis. In the present study the genomic sequence was obtained from retroviral transfected mouse showing multiple tumors…

Introduction


Gene therapy, integrating vectors carrying therapeutic transgene sequences offers the potential for a permanent cure of genetic diseases by stable vector insertion into the patients chromosomes (1). However there are some reports indicating occurrence of tumors at later stage in transgenic animal and that's why it is important to know probability of non specific integration of this transgene and its effect on cellular homeostasis. As per the present understanding the integration is semi-random in nature and having partial preference towards sequences in or near the coding regions of expressed genes (1). Integration in these places may lead to up or down regulation of that particular gene and hence increase the probability of interference in cellular homeostasis. Based on above observations, it is highly recommended to verify the insertion loci of given vectors in model system. Based on bio-informatical analysis of given sequence, we were able to demonstrated that Viral vector integrates in vicinity to gene called Nfib (Nuclear factor I/B) and interferes with it normal functioning. Detailed investigation and database search indicates Nfib has potential role in cell cycle regulation and oncogenesis.
Vectors, transfection, cloning, amplification and sequencing were performed as per previously mention protocol (1). ...
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