AAV-mediated gene targeting is highly dependent on the host cell-cycle status, requiring DNA synthesis for efficient targeting.
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Cell-cycle dependence may constrain AAV gene targeting in quiescent tissues; leaves open whether proliferation modulation can improve efficiency in vivo.
Adeno-associated virus (AAV) vectors can be used to introduce site-specific mutations into homologous chromosomal sequences. There are many potential applications of this technique, but the process of AAV-mediated gene targeting and factors that influence targeting efficiency are not completely understood. We investigated the dependence of AAV-mediated gene targeting on the host cell-cycle status. The frequency of gene targeting by AAV vectors was compared in dividing and serum-arrested normal human fibroblast cultures. Gene targeting occurred in arrested fibroblast cultures at 0.15 to 1.1% the frequency of dividing cultures, and only took place in cells that had undergone DNA synthesis. Gene targeting was also reduced when DNA synthesis was inhibited by hydroxyurea.
Trobridge et al. (2005) studied this question.
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