A model for the mechanism of precise integration of a microinjected transgene
A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DN...
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| Veröffentlicht in: | Transgenic research 1996-05, Vol.5 (3), p.171-177 |
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| container_title | Transgenic research |
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| creator | MCFARLANE, M WILSON, J. B |
| description | A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DNA sequence of the transgene and cellular junctions have been determined. A comparison between wild type and transgenic mutant sequences at the site of insertion revealed that there was no loss or rearrangement of cellular DNA upon integration of the transgene. The cellular sequences at the transgene 5' and 3' joins are contiguous in the wild type. The integrant exists as a head to tail tandem dimer with minimal loss of sequence compared with the injected monomer. Analysis of the site of insertion has revealed a 5 bp homology between the 5' end of the transgene and the cellular sequences. In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites of DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, the Escherichia coli Chi site and the meiotic recombination hotspot within the E beta gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed. |
| doi_str_mv | 10.1007/BF01969706 |
| format | Article |
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In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites of DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, the Escherichia coli Chi site and the meiotic recombination hotspot within the E beta gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed.</description><identifier>ISSN: 0962-8819</identifier><identifier>EISSN: 1573-9368</identifier><identifier>DOI: 10.1007/BF01969706</identifier><identifier>PMID: 8673144</identifier><language>eng</language><publisher>Dordrecht: Springer</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Cleft Palate - genetics ; DNA - genetics ; DNA Topoisomerases, Type I - metabolism ; DNA, Recombinant - genetics ; Epidermis - pathology ; Female ; Fundamental and applied biological sciences. Psychology ; Gene Conversion ; Genes, Viral ; Genetic engineering ; Genetic technics ; Genic rearrangement. Recombination. Transposable element ; Herpesvirus 4, Human - genetics ; Humans ; Hyperplasia ; Methods. Procedures. Technologies ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic - genetics ; Microinjections ; Models, Genetic ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Polyomavirus - genetics ; Recombination, Genetic ; Regulatory Sequences, Nucleic Acid ; Sequence Homology, Nucleic Acid ; Transgenes ; Transgenic animals ; Transgenic animals and transgenic plants ; Viral Matrix Proteins - biosynthesis ; Viral Matrix Proteins - genetics ; X Chromosome - genetics</subject><ispartof>Transgenic research, 1996-05, Vol.5 (3), p.171-177</ispartof><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c342t-2aefc75e329fd04d1b0a8ef92b1ec2c50fa6981f20ced61d70efa3edc73ab4983</citedby><cites>FETCH-LOGICAL-c342t-2aefc75e329fd04d1b0a8ef92b1ec2c50fa6981f20ced61d70efa3edc73ab4983</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3079438$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8673144$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>MCFARLANE, M</creatorcontrib><creatorcontrib>WILSON, J. B</creatorcontrib><title>A model for the mechanism of precise integration of a microinjected transgene</title><title>Transgenic research</title><addtitle>Transgenic Res</addtitle><description>A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DNA sequence of the transgene and cellular junctions have been determined. A comparison between wild type and transgenic mutant sequences at the site of insertion revealed that there was no loss or rearrangement of cellular DNA upon integration of the transgene. The cellular sequences at the transgene 5' and 3' joins are contiguous in the wild type. The integrant exists as a head to tail tandem dimer with minimal loss of sequence compared with the injected monomer. Analysis of the site of insertion has revealed a 5 bp homology between the 5' end of the transgene and the cellular sequences. In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites of DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, the Escherichia coli Chi site and the meiotic recombination hotspot within the E beta gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cleft Palate - genetics</subject><subject>DNA - genetics</subject><subject>DNA Topoisomerases, Type I - metabolism</subject><subject>DNA, Recombinant - genetics</subject><subject>Epidermis - pathology</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Conversion</subject><subject>Genes, Viral</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genic rearrangement. Recombination. Transposable element</subject><subject>Herpesvirus 4, Human - genetics</subject><subject>Humans</subject><subject>Hyperplasia</subject><subject>Methods. Procedures. Technologies</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Transgenic - genetics</subject><subject>Microinjections</subject><subject>Models, Genetic</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Polyomavirus - genetics</subject><subject>Recombination, Genetic</subject><subject>Regulatory Sequences, Nucleic Acid</subject><subject>Sequence Homology, Nucleic Acid</subject><subject>Transgenes</subject><subject>Transgenic animals</subject><subject>Transgenic animals and transgenic plants</subject><subject>Viral Matrix Proteins - biosynthesis</subject><subject>Viral Matrix Proteins - genetics</subject><subject>X Chromosome - genetics</subject><issn>0962-8819</issn><issn>1573-9368</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtLAzEUhYMotVY37oUsxIUwmsdMHstarAoVN7oeMslNmzKPmkwX_nuntNSlqwvnfBy4H0LXlDxQQuTj05xQLbQk4gSNaSF5prlQp2hMtGCZUlSfo4uU1oQMuOIjNFJCcprnY_Q-xU3noMa-i7hfAW7ArkwbUoM7jzcRbEiAQ9vDMpo-dO0uNrgJNnahXYPtweE-mjYtoYVLdOZNneDqcCfoa_78OXvNFh8vb7PpIrM8Z33GDHgrC-BMe0dyRytiFHjNKgqW2YJ4I7SinhELTlAnCXjDwVnJTZVrxSfobr-7id33FlJfNiFZqGvTQrdNpVQkF1IV_4K0EFTSgg3g_R4c_kopgi83MTQm_pSUlDvJ5Z_kAb45rG6rBtwRPVgd-ttDb5I1tR_0DBqPGCdS51zxXztmg9M</recordid><startdate>19960501</startdate><enddate>19960501</enddate><creator>MCFARLANE, M</creator><creator>WILSON, J. B</creator><general>Springer</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19960501</creationdate><title>A model for the mechanism of precise integration of a microinjected transgene</title><author>MCFARLANE, M ; WILSON, J. B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c342t-2aefc75e329fd04d1b0a8ef92b1ec2c50fa6981f20ced61d70efa3edc73ab4983</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cleft Palate - genetics</topic><topic>DNA - genetics</topic><topic>DNA Topoisomerases, Type I - metabolism</topic><topic>DNA, Recombinant - genetics</topic><topic>Epidermis - pathology</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Conversion</topic><topic>Genes, Viral</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genic rearrangement. Recombination. Transposable element</topic><topic>Herpesvirus 4, Human - genetics</topic><topic>Humans</topic><topic>Hyperplasia</topic><topic>Methods. Procedures. Technologies</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Transgenic - genetics</topic><topic>Microinjections</topic><topic>Models, Genetic</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Polyomavirus - genetics</topic><topic>Recombination, Genetic</topic><topic>Regulatory Sequences, Nucleic Acid</topic><topic>Sequence Homology, Nucleic Acid</topic><topic>Transgenes</topic><topic>Transgenic animals</topic><topic>Transgenic animals and transgenic plants</topic><topic>Viral Matrix Proteins - biosynthesis</topic><topic>Viral Matrix Proteins - genetics</topic><topic>X Chromosome - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>MCFARLANE, M</creatorcontrib><creatorcontrib>WILSON, J. B</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Transgenic research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>MCFARLANE, M</au><au>WILSON, J. B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A model for the mechanism of precise integration of a microinjected transgene</atitle><jtitle>Transgenic research</jtitle><addtitle>Transgenic Res</addtitle><date>1996-05-01</date><risdate>1996</risdate><volume>5</volume><issue>3</issue><spage>171</spage><epage>177</epage><pages>171-177</pages><issn>0962-8819</issn><eissn>1573-9368</eissn><abstract>A unique transgenic mouse line has undergone transgene integration in a very precise fashion. The phenotype displayed by mice of the line followed the predicted inheritance patterns for X-linked transgene insertion which has been confirmed. In order to investigate the mechanism of integration the DNA sequence of the transgene and cellular junctions have been determined. A comparison between wild type and transgenic mutant sequences at the site of insertion revealed that there was no loss or rearrangement of cellular DNA upon integration of the transgene. The cellular sequences at the transgene 5' and 3' joins are contiguous in the wild type. The integrant exists as a head to tail tandem dimer with minimal loss of sequence compared with the injected monomer. Analysis of the site of insertion has revealed a 5 bp homology between the 5' end of the transgene and the cellular sequences. In addition, adjacent to the site of insertion within the cellular sequences, there are several sequence motifs implicated in recombination events including a clustering of strong consensus sites of DNA topoisomerase type I and a region of homology to the human minisatellite consensus core sequence, the Escherichia coli Chi site and the meiotic recombination hotspot within the E beta gene of the murine major histocompatibility complex. This clustering of features is likely to have been factorial in the integrity of the insertion event. A model depicting the mechanism of this precise integration is proposed.</abstract><cop>Dordrecht</cop><pub>Springer</pub><pmid>8673144</pmid><doi>10.1007/BF01969706</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Base Sequence Biological and medical sciences Biotechnology Cleft Palate - genetics DNA - genetics DNA Topoisomerases, Type I - metabolism DNA, Recombinant - genetics Epidermis - pathology Female Fundamental and applied biological sciences. Psychology Gene Conversion Genes, Viral Genetic engineering Genetic technics Genic rearrangement. Recombination. Transposable element Herpesvirus 4, Human - genetics Humans Hyperplasia Methods. Procedures. Technologies Mice Mice, Inbred C57BL Mice, Transgenic - genetics Microinjections Models, Genetic Molecular and cellular biology Molecular genetics Molecular Sequence Data Polyomavirus - genetics Recombination, Genetic Regulatory Sequences, Nucleic Acid Sequence Homology, Nucleic Acid Transgenes Transgenic animals Transgenic animals and transgenic plants Viral Matrix Proteins - biosynthesis Viral Matrix Proteins - genetics X Chromosome - genetics |
| title | A model for the mechanism of precise integration of a microinjected transgene |
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