Characterization of Cre Expression in Bac-Pcp2-Ires-Cre Transgenic Mice

This dissertation, "Characterization of Cre Expression in BAC-Pcp2-IRES-Cre Transgenic Mice" by Hoi-lam, Alam, Ng, 吳凱琳, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled Characterization of Cre expression in BAC-Pcp2-IRES-Cre Transgenic mice submitted by Ng Hoi Lam Alam for the degree of Master of Philosophy at the University of Hong Kong in August 2005 It has long been known that intracellular transportation system is relied on the cooperation and interdependence of microfilaments and microtubule networks (Schliwa, M., 1999). Previous studies by Huang et al. suggest that the motors associated with these two cytoskeletal networks are coordinated and interacted directly through an actin-based motor, MyoVA and a microtubule-based motor, the ubiquitously expressed kinesin heavy chain (KhcU or kinesin-1) (Huang, J.D. et al., 1999). In order to study the role of kinesin-1, PC-specific KhcU knockout mice will be generated. This relies on: 1) mouse with loxP sites inserted in KhcU gene and 2) mouse expresses cre activity in cerebellar PCs. Using BAC as a carrier, cre cDNA is inserted after the promoter of Pcp2 and 1introduced into the genome to generate a transgenic mouse line BAC-Pcp2-IRES-Cre (Zhang et al., 2004). Under the control of Pcp2 promoter, Cre activity should be expressed only in cerebellar PC and retinal rod bipolar cells. Using ROSA26 and Z/EG reporter mouse lines, Cre activities are detected in different postnatal days. My results show that Cre activity starts at postnatal day 6 and 7 in cerebellar PC and retinal rod bipolar cells, respectively. This transgenic mouse line shows high specificity that Cre expression is detected only in cerebellar PC and retinal rod bipolar cells, but not in other parts of the body. In addition, there is no observable difference in the morphology and developmental pattern of these cells. As a result, BAC-Pcp2-IRES-Cre can be applied to generate PC- or rod bipolar cell-specific kinesin-1 knockout mice. 2 DOI: 10.5353/th_b3627824 Subjects: Genetic recombination Gene expression Kinesin Biological transport Transgenic mice