Research On The Role Of Sry-box₂ Gene Sox 2 In Rats Liver Regeneration

The member of the SOX gene family SRY-box 2 sox₂ is related to development and differentiation of cell. Effects of sox₂ on the process of liver regeneration in rats were studied by use the technique of hydrodynamics-based transgene, via the method of gene addition and RNA interference in this paper, in order to provide experimental basis for clarifying the function of sox₂ and the molecular mechanism of liver regeneration.sox₂ was cloned according to the published information, two target sites on the design two interfering RNA fragments were selected based on the gene sequence, expressive and interference vectors were constructed linked with the right fragments designed above: the expressive vector was pEGFP-N1- sox₂, and the other two interference vector were pGenesil-1.0-s199, pGenesil-1.0-s370. Then the test vectors pGenesil-1.0-s199-sox₂, pGenesil-1.0-s370-sox₂ were constructed linked with the ORFof sox₂, which was double digested from the expressive vector. The constructed test vectors were transgened based on hydrodynamics, in 0, 6, 12, 24, 36, 48, 60, 72 and 84h after transgene the interventionist fragments were determined whether the role of the target site by observing the interventionist test vectors rate of green fluorescent protein positive cells (transfection rate) compared with the control there is an obvious decrease in the intervention designed. The best transgenic time after partial hepatectomy was obtained by transfection of vector via hydrodynamics expression in rat liver tissue the peak point in time, combined with sox₂ expression in rat liver tissue the peak point in time. The liver materials obtained 72, 120, and 168h after partial hepatectomy were observed and analyzed by using fluorescence microscopy techniques, statistical methods and conventional histological technique on morphology structure, expression kinetics and index of liver regeneration, etc.The tests were showed: cloned sox₂ sequence is correct, two fragments were interfering with the more pronounced interference effect, the vector was successfully constructed. The best transgenic time was in 0h after partial hepatectomy. Transfer interference vector pGenesil-1.0-s370, the index of liver regeneration in liver has some changes compared with the control.