Williams, AJ; (2006) Studies on aspects that affect chromatin structure and function of the human CD2 locus control region. Doctoral thesis , UCL (University College London).

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Abstract

The overall aim of this project is to study the control of hCD2 gene expression and how different parts of the gene contribute to its regulation. This was achieved by searching for novel regulatory elements within the hCD2 gene whose function could be addressed by deleting them using an in vivo Cre/loxP deletion strategy. For this purpose it was necessary to develop tools that would, in future, enable the in vivo tissue-specific deletion of these identified sequences utilising this approach. To identify novel regulatory elements within the hCD2 gene a number of different methods were used. Previously intron 4 has been shown to confer elevated levels of expression on hCD2 transgenes, however its function has never been studied in the context of variegation. Therefore, a construct including intron 4 and the truncated LCR was generated. This vector allowed elevated levels of hCD2 expression without conferring protection against PEV in transgenic mice, thus confirming that intron 4 could not compensate for a debilitated LCR. In addition, it was shown that decreasing transgene copy number whilst retaining the same site of integration affected hCD2 variegation in a manner that was dependant upon the site of integration. Therefore, we established that the inclusion of intron 4 in transgenes would be used for variegation studies whilst ensuring high levels of expression.

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