The human pre-B cell line Nalm-6 is highly proficient in gene targeting by homologous recombination.
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.
Adachi
Noritaka
N
Kihara Institute for Biological Research, Graduate School of Integrated Science, Yokohama City University, Japan. nadachi@yokohama-cu.ac.jp
So
Sairei
S
Iiizumi
Susumu
S
Nomura
Yuji
Y
Murai
Kyoko
K
Yamakawa
Chie
C
Miyagawa
Kiyoshi
K
Koyama
Hideki
H
D001402:B-Lymphocytes / Q000378:metabolism*; D002460:Cell Line / Q000378:metabolism; D045744:Cell Line, Tumor; D002869:Chromosome Aberrations; D018390:Gene Targeting; D016158:Genes, p53; D006801:Humans; D051718:MutS Homolog 2 Protein / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic
C497172:MSH2 protein, human; D051718:MutS Homolog 2 Protein