Research Article
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Published Online: 11 January 2006

The Human Pre-B Cell Line Nalm-6 Is Highly Proficient in Gene Targeting by Homologous Recombination

Publication: DNA and Cell Biology
Volume 25, Issue Number 1

Abstract

Gene targeting provides a powerful means for analyzing gene function, as exemplified by knockout mouse studies and recent work with the highly recombinogenic chicken DT40 B-lymphocyte line. In human cultured cells, however, the low frequency of gene targeting is a serious barrier to efficiently generate knockout clones. Moreover, commonly used human cell lines are karyotypically abnormal or unstable. Here, we show using promoterless targeting constructs that Nalm-6, a human pre-B ALL cell line, is highly proficient for gene targeting by homologous recombination. Indeed, the efficiency of TP53 gene targeting in Nalm-6 appears nearly two orders of magnitude higher than that in HCT116, a colon cancer cell line popularly used for gene targeting. Expression analysis revealed a lack of MSH2 expression in this cell line. As Nalm-6 has a stable neardiploid karyotype with normal p53 status, our results underscore the usefulness of Nalm-6 for gene knockout studies in humans.

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Published In

cover image DNA and Cell Biology
DNA and Cell Biology
Volume 25Issue Number 1January 2006
Pages: 19 - 24
PubMed: 16405397

History

Published online: 11 January 2006
Published in print: January 2006

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Noritaka Adachi
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Sairei So
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Susumu Iiizumi
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Yuji Nomura
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Kyoko Murai
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Chie Yamakawa
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.
Kiyoshi Miyagawa
Department of Radiation Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
Hideki Koyama
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Yokohama, Japan.

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