Functional Analysis of proteins involved in chromosomal translocations in hematological neoplasms

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Translin has been identified as a DNA binding protein that specifically recognizes the consensus sequences motifs at breakpoint junctions in many chromosomal translocations in human lymphoid neoplasms. In the past analysis, many functional features of Translin protein have been characterized. However the physiological function and the association to chromasomal translocations have not been clarified until now. To resolve the problem, we planned the project to make the KO mouse of translin and the associated protein, RP58. Using the known genomic structures and sequences of the both genes, we constructed the target vectors to remove the functional portions of the genes. After the electoroporations of the target vectors to ES cells, we selected the candidate clones using the suitable medium for selection. The genomic DNA of the survived clones were examined by the southern blotting analysis and the objective ES clones were identified. Using these clones, we have finally succeed to make the KO mouse of Translin and RP58 by the conventional method. Now we are energetically making analysis to clarified the physiological functions of both proteins, and would like to prove their involvement to chromasomal translocations.