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Conditional deletion of S6 ribosomal protein gene by CD4 Cre transgene

To study the relationship between cell growth and cell cycle progression, we interfered with the ribosomal biogenesis by deleting one allele of S6 ribosomal protein gene in thymus by using Cre recombinase under the control of CD4 promoter. In that model we studied thymocyte development and funtional properties of mature T cells. Cre/lox P system was used to delete one S6 allele in double positive cells in thymus. Pups from crossings of homozygous floxed S6 mice with CD4 Cre mice were screened for Cre transgene with PCR. Deletion of one S6 allele was confirmed by Southern blot analysis and expression of S6 mRNA was measured by Northern blot analysis. Flow cytometry was used to study population of T cells during development and cell size was measured by FSC. Proliferative capacity and cytokine production were analyzed to estimate function of peripheral T cells.Conditional deletion of one S6 allele did not have any consequence on differentiation, size and number of thymocytes, although deletion was 100% effective. However, we observed dramatic effect on number of mature T cells in spleen, whose size was normal. Our goal is to determine the effect of limited ribosome biogenesis on cell growth and cell cycle progression and differentiation of T cells. This study can help us to better understand pathological processes caracterized by impaired ribosome production.

S6 ribosomal protein; CD4 Cre transgene; T lymphocytes

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