In vivo site-specific recombination using the beta-rec/six system.

The prokaryotic beta serine recombinase (beta-rec) catalyzes site-specific recombination between two directly oriented six sites (93 bp) in mammalian cells, both in episomal and in chromosomally integrated substrates. The beta-rec/six exclusive intramolecular site-specific recombination (SSR) system has been proposed as a suitable approach when several independently controlled recombination events are needed in a single cell. Here we explored the use of the beta-rec/six system for selective induction of genome-targeted modifications. We generated and analyzed mouse transgenic lines (Tgbeta) expressing beta-rec under the control of the Lck promoter. beta-rec activity was demonstrated, and there was no evidence of alterations to thymic or peripheral T cell development. We developed two transgenic mouse lines harboring different target sequences (Tgrec and KOsix) and analyzed the effect of beta-rec expression on these animals. The results indicate that the beta-rec/six SSR system is functional for in vivo gene-targeting applications.

[1]  C. Stemmer,et al.  Functionality of the β/six Site-Specific Recombination System in Tobacco and Arabidopsis: A Novel Tool for Genetic Engineering of Plant Genomes , 2007, Plant Molecular Biology.

[2]  S. Murray,et al.  Generation of a Snail1 (Snai1) conditional null allele , 2006, Genesis.

[3]  D. Sorrell,et al.  Targeted modification of mammalian genomes. , 2005, Biotechnology advances.

[4]  S. Woo,et al.  Complete and persistent phenotypic correction of phenylketonuria in mice by site-specific genome integration of murine phenylalanine hydroxylase cDNA , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[5]  P. Rice.  Resolving integral questions in site-specific recombination , 2005, Nature Structural &Molecular Biology.

[6]  Philippe Soriano,et al.  Cryptic boundaries in roof plate and choroid plexus identified by intersectional gene activation , 2003, Nature Genetics.

[7]  Margaret C. M. Smith,et al.  Diversity in the serine recombinases , 2002, Molecular microbiology.

[8]  M. Lewandoski.  Conditional control of gene expression in the mouse , 2001, Nature Reviews Genetics.

[9]  A. Berns,et al.  Growth inhibition and DNA damage induced by Cre recombinase in mammalian cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Manuel A. González,et al.  New Insights into Host Factor Requirements for Prokaryotic β-Recombinase-mediated Reactions in Mammalian Cells* , 2001, The Journal of Biological Chemistry.

[11]  M. Capecchi,et al.  Illegitimate Cre-dependent chromosome rearrangements in transgenic mouse spermatids. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  E. Zubko,et al.  Intrachromosomal recombination between attP regions as a tool to remove selectable marker genes from tobacco transgenes , 2000, Nature Biotechnology.

[13]  B. Thyagarajan,et al.  Mammalian genomes contain active recombinase recognition sites. , 2000, Gene.

[14]  Wen-Hwa Lee,et al.  Temporal, spatial, and cell type–specific control of Cre-mediated DNA recombination in transgenic mice , 1999, Nature Biotechnology.

[15]  D. Metzger,et al.  Engineering the mouse genome by site-specific recombination. , 1999, Current opinion in biotechnology.

[16]  C. Martínez-A,et al.  The Prokaryotic β-Recombinase Catalyzes Site-specific Recombination in Mammalian Cells* , 1999, The Journal of Biological Chemistry.

[17]  B. Chang,et al.  Liver-specific Inactivation of the Abetalipoproteinemia Gene Completely Abrogates Very Low Density Lipoprotein/Low Density Lipoprotein Production in a Viable Conditional Knockout Mouse* , 1999, The Journal of Biological Chemistry.

[18]  J. Rossant,et al.  "Cre"-ating mouse mutants-a meeting review on conditional mouse genetics. , 1999, Genes & development.

[19]  C. Blancher,et al.  Retinal dysplasia in mice lacking p56lck , 1998, Oncogene.

[20]  G. Martin,et al.  An Fgf8 mutant allelic series generated by Cre- and Flp-mediated recombination , 1998, Nature Genetics.

[21]  B. Sauer,et al.  Brief expression of a GFP cre fusion gene in embryonic stem cells allows rapid retrieval of site-specific genomic deletions. , 1997, Nucleic acids research.

[22]  B. A. French,et al.  Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo. , 1997, The Journal of clinical investigation.

[23]  B. Spiegelman,et al.  Targeted expression of Cre recombinase to adipose tissue of transgenic mice directs adipose-specific excision of loxP-flanked gene segments. , 1997, Nucleic acids research.

[24]  F. Watrin,et al.  Deletion of the mouse T-cell receptor beta gene enhancer blocks alphabeta T-cell development. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[25]  F. Rojo,et al.  Site-specific recombination by the beta protein from the streptococcal plasmid pSM19035: minimal recombination sequences and crossing over site. , 1996, Nucleic acids research.

[26]  S. Dymecki.  Flp recombinase promotes site-specific DNA recombination in embryonic stem cells and transgenic mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[27]  C. Gutiérrez,et al.  The role of the chromatin‐associated protein Hbsu in β‐mediated DNA recombination is to facilitate the joining of distant recombination sites , 1995, Molecular microbiology.

[28]  F. Rojo,et al.  The beta recombinase of plasmid pSM19035 binds to two adjacent sites, making different contacts at each of them. , 1995, Nucleic acids research.

[29]  F. Rojo,et al.  The Bacillussubtilis Histone-like Protein Hbsu Is Required for DNA Resolution and DNA Inversion Mediated by the Recombinase of Plasmid pSM19035 (*) , 1995, The Journal of Biological Chemistry.

[30]  B. Sauer.  Site-specific recombination: developments and applications. , 1994, Current opinion in biotechnology.

[31]  K. Rajewsky,et al.  Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. , 1994, Science.

[32]  F. Rojo,et al.  A novel site-specific recombinase encoded by the Streptococcus pyogenes plasmid pSM19035. , 1994, Journal of molecular biology.

[33]  P. Stanley,et al.  Mice lacking N-acetylglucosaminyltransferase I activity die at mid-gestation, revealing an essential role for complex or hybrid N-linked carbohydrates. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[34]  F. Rojo,et al.  Purification of the beta product encoded by the Streptococcus pyogenes plasmid pSM19035. A putative DNA recombinase required to resolve plasmid oligomers. , 1993, FEBS letters.

[35]  G. Sclar,et al.  Thymic overexpression of Ttg-1 in transgenic mice results in T-cell acute lymphoblastic leukemia/lymphoma , 1992, Molecular and cellular biology.

[36]  J. Marth,et al.  Tissue- and site-specific DNA recombination in transgenic mice. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[37]  I. Horak,et al.  Development and function of T cells in mice rendered interleukin-2 deficient by gene targeting , 1991, Nature.

[38]  N. Henderson,et al.  Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[39]  B. Sauer.  Manipulation of transgenes by site-specific recombination: use of Cre recombinase. , 1993, Methods in enzymology.