Specific mutations induced by triplex-forming oligonucleotides in mice.

Triplex-forming oligonucleotides (TFOs) recognize and bind to specific duplex DNA sequences and have been used extensively to modify gene function in cells. Although germ line mutations can be incorporated by means of embryonic stem cell technology, little progress has been made toward introducing mutations in somatic cells of living organisms. Here we demonstrate that TFOs can induce mutations at specific genomic sites in somatic cells of adult mice. Mutation detection was facilitated by the use of transgenic mice bearing chromosomal copies of the supF and cII reporter genes. Mice treated with a supF-targeted TFO displayed about fivefold greater mutation frequencies in the supF gene compared with mice treated with a scrambled sequence control oligomer. No mutagenesis was detected in the control gene (cII) with either oligonucleotide. These results demonstrate that site-specific, TFO-directed genome modification can be accomplished in intact animals.

[1]  E. Luzi,et al.  Designing ribozymes for the inhibition of gene expression. , 1998, Trends in biotechnology.

[2]  P. Glazer,et al.  Elevated levels of mutation in multiple tissues of mice deficient in the DNA mismatch repair gene Pms2. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[3]  P. Nielsen,et al.  Progress in developing PNA as a gene-targeted drug. , 1997, Antisense & nucleic acid drug development.

[4]  P. Glazer,et al.  Targeted mutagenesis in mammalian cells mediated by intracellular triple helix formation , 1995, Molecular and cellular biology.

[5]  P. Glazer,et al.  Chromosomal mutations induced by triplex-forming oligonucleotides in mammalian cells. , 1999, Nucleic acids research.

[6]  T. Prolla,et al.  Different mutator phenotypes in Mlh1- versus Pms2-deficient mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[7]  B. Myhr.  Validation studies with Muta™ mouse: A transgenic mouse model for detecting mutations in vivo , 1991, Environmental and molecular mutagenesis.

[8]  M. Inouye,et al.  Frameshift mutations and the genetic code. This paper is dedicated to Professor Theodosius Dobzhansky on the occasion of his 66th birthday. , 1966, Cold Spring Harbor symposia on quantitative biology.

[9]  S. Agrawal,et al.  Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Tinsley,et al.  In vivo stability and kinetics of absorption and disposition of 3' phosphopropyl amine oligonucleotides. , 1992, Nucleic acids research.

[11]  A. Harel-Bellan,et al.  Unambiguous demonstration of triple-helix-directed gene modification. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  J. H. Wilson,et al.  Triplex-directed modification of genes and gene activity. , 1998, Trends in biochemical sciences.

[13]  U. Galderisi,et al.  Antisense oligonucleotides as therapeutic agents , 1999, Journal of cellular physiology.

[14]  M. Famulok,et al.  Oligonucleotide aptamers that recognize small molecules. , 1999, Current opinion in structural biology.

[15]  P. Glazer,et al.  Targeted gene knockout mediated by triple helix forming oligonucleotides , 1998, Nature Genetics.

[16]  P. Miller,et al.  Disposition and metabolism of oligodeoxynucleoside methylphosphonate following a single i.v. injection in mice. , 1990, Drug metabolism and disposition: the biological fate of chemicals.

[17]  Peter B. Dervan,et al.  Regulation of gene expression by small molecules , 1997, Nature.

[18]  D. Gordenin,et al.  Yeast ARMs (DNA at-risk motifs) can reveal sources of genome instability. , 1998, Mutation research.

[19]  M. Rice,et al.  Correction of the Mutation Responsible for Sickle Cell Anemia by an RNA-DNA Oligonucleotide , 1996, Science.

[20]  J. French,et al.  Analysis of genetic instability during mammary tumor progression using a novel selection-based assay for in vivo mutations in a bacteriophage lambda transgene target. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[21]  P. Glazer,et al.  Triple-Helix Formation Induces Recombination in Mammalian Cells via a Nucleotide Excision Repair-Dependent Pathway , 2000, Molecular and Cellular Biology.

[22]  P. Hanawalt.  Transcription-coupled repair and human disease. , 1994, Science.

[23]  P. Glazer,et al.  Mutagenesis in Mammalian Cells Induced by Triple Helix Formation and Transcription-Coupled Repair , 1996, Science.