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TALENs Construction: Slowly but Surely

  • Hegazy, Wael Abdel Halim (Department of Microbiology and Immunology, Faculty of Pharmacy, Zagzig University) ;
  • Youns, Mahmoud (Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, Helwan University)
  • Published : 2016.07.01

Abstract

Cancer is thought to be a direct result of transcriptional misregulation. Broad analysis of transcriptional regulatory elements in healthy and cancer cells is needed to understand cancer development. Nucleases regulatory domains are recruited to bind and manipulate a specific genomic locus with high efficacy and specificity. TALENs (transcription activator-like effector nuclease) fused to endonuclease FokI have been used widely to target specific sequences to edit several genes in healthy and cancer cells. This approach is promising to target specific cancer genes and for this purpose it is needed to pack such TALENs into viral vectors. There are some considerations which control the success of this approach, targeting appropriate sequences with efficient construction of TALENs being crucial factors. We face some obstacles in construction of TALENs; in this study we made a modification to the method of Cermk et al 2011 and added one step to make it easier and increase the availability of constructs.

Keywords

TALENs;RVDs;Golden gate construction;cancer therapy

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