Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model

Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifi...
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Autor*in:	Charizopoulou, Nikoletta [verfasserIn] Jansen, Silke Dorsch, Martina Stanke, Frauke Dorin, Julia R Hedrich, Hans-Jürgen Tümmler, Burkhard

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2004

Schlagwörter:	Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Congenic Strain Cftr Gene Excision Event

Anmerkung:	© Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

Übergeordnetes Werk:	Enthalten in: BMC genetics - London : BioMed Central, 2000, 5(2004), 1 vom: 21. Apr.
Übergeordnetes Werk:	volume:5 ; year:2004 ; number:1 ; day:21 ; month:04

Links:	Volltext

DOI / URN:	10.1186/1471-2156-5-6

Katalog-ID:	SPR026999366

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520			\|a Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice.
650		4	\|a Cystic Fibrosis \|7 (dpeaa)DE-He213
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700	1		\|a Hedrich, Hans-Jürgen \|4 aut
700	1		\|a Tümmler, Burkhard \|4 aut
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publishDate	2004
allfields	10.1186/1471-2156-5-6 doi (DE-627)SPR026999366 (SPR)1471-2156-5-6-e DE-627 ger DE-627 rakwb eng Charizopoulou, Nikoletta verfasserin aut Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model 2004 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213 Jansen, Silke aut Dorsch, Martina aut Stanke, Frauke aut Dorin, Julia R aut Hedrich, Hans-Jürgen aut Tümmler, Burkhard aut Enthalten in BMC genetics London : BioMed Central, 2000 5(2004), 1 vom: 21. Apr. (DE-627)326644938 (DE-600)2041497-3 1471-2156 nnns volume:5 year:2004 number:1 day:21 month:04 https://dx.doi.org/10.1186/1471-2156-5-6 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 5 2004 1 21 04
spelling	10.1186/1471-2156-5-6 doi (DE-627)SPR026999366 (SPR)1471-2156-5-6-e DE-627 ger DE-627 rakwb eng Charizopoulou, Nikoletta verfasserin aut Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model 2004 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213 Jansen, Silke aut Dorsch, Martina aut Stanke, Frauke aut Dorin, Julia R aut Hedrich, Hans-Jürgen aut Tümmler, Burkhard aut Enthalten in BMC genetics London : BioMed Central, 2000 5(2004), 1 vom: 21. Apr. (DE-627)326644938 (DE-600)2041497-3 1471-2156 nnns volume:5 year:2004 number:1 day:21 month:04 https://dx.doi.org/10.1186/1471-2156-5-6 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 5 2004 1 21 04
allfields_unstemmed	10.1186/1471-2156-5-6 doi (DE-627)SPR026999366 (SPR)1471-2156-5-6-e DE-627 ger DE-627 rakwb eng Charizopoulou, Nikoletta verfasserin aut Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model 2004 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213 Jansen, Silke aut Dorsch, Martina aut Stanke, Frauke aut Dorin, Julia R aut Hedrich, Hans-Jürgen aut Tümmler, Burkhard aut Enthalten in BMC genetics London : BioMed Central, 2000 5(2004), 1 vom: 21. Apr. (DE-627)326644938 (DE-600)2041497-3 1471-2156 nnns volume:5 year:2004 number:1 day:21 month:04 https://dx.doi.org/10.1186/1471-2156-5-6 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 5 2004 1 21 04
allfieldsGer	10.1186/1471-2156-5-6 doi (DE-627)SPR026999366 (SPR)1471-2156-5-6-e DE-627 ger DE-627 rakwb eng Charizopoulou, Nikoletta verfasserin aut Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model 2004 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213 Jansen, Silke aut Dorsch, Martina aut Stanke, Frauke aut Dorin, Julia R aut Hedrich, Hans-Jürgen aut Tümmler, Burkhard aut Enthalten in BMC genetics London : BioMed Central, 2000 5(2004), 1 vom: 21. Apr. (DE-627)326644938 (DE-600)2041497-3 1471-2156 nnns volume:5 year:2004 number:1 day:21 month:04 https://dx.doi.org/10.1186/1471-2156-5-6 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 5 2004 1 21 04
allfieldsSound	10.1186/1471-2156-5-6 doi (DE-627)SPR026999366 (SPR)1471-2156-5-6-e DE-627 ger DE-627 rakwb eng Charizopoulou, Nikoletta verfasserin aut Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model 2004 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213 Jansen, Silke aut Dorsch, Martina aut Stanke, Frauke aut Dorin, Julia R aut Hedrich, Hans-Jürgen aut Tümmler, Burkhard aut Enthalten in BMC genetics London : BioMed Central, 2000 5(2004), 1 vom: 21. Apr. (DE-627)326644938 (DE-600)2041497-3 1471-2156 nnns volume:5 year:2004 number:1 day:21 month:04 https://dx.doi.org/10.1186/1471-2156-5-6 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 5 2004 1 21 04
language	English
source	Enthalten in BMC genetics 5(2004), 1 vom: 21. Apr. volume:5 year:2004 number:1 day:21 month:04
sourceStr	Enthalten in BMC genetics 5(2004), 1 vom: 21. Apr. volume:5 year:2004 number:1 day:21 month:04
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Congenic Strain Cftr Gene Excision Event
isfreeaccess_bool	true
container_title	BMC genetics
authorswithroles_txt_mv	Charizopoulou, Nikoletta @@aut@@ Jansen, Silke @@aut@@ Dorsch, Martina @@aut@@ Stanke, Frauke @@aut@@ Dorin, Julia R @@aut@@ Hedrich, Hans-Jürgen @@aut@@ Tümmler, Burkhard @@aut@@
publishDateDaySort_date	2004-04-21T00:00:00Z
hierarchy_top_id	326644938
id	SPR026999366
language_de	englisch
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This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. 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author	Charizopoulou, Nikoletta
spellingShingle	Charizopoulou, Nikoletta misc Cystic Fibrosis misc Cystic Fibrosis Transmembrane Conductance Regulator misc Congenic Strain misc Cftr Gene misc Excision Event Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model
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topic_title	Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model Cystic Fibrosis (dpeaa)DE-He213 Cystic Fibrosis Transmembrane Conductance Regulator (dpeaa)DE-He213 Congenic Strain (dpeaa)DE-He213 Cftr Gene (dpeaa)DE-He213 Excision Event (dpeaa)DE-He213
topic	misc Cystic Fibrosis misc Cystic Fibrosis Transmembrane Conductance Regulator misc Congenic Strain misc Cftr Gene misc Excision Event
topic_unstemmed	misc Cystic Fibrosis misc Cystic Fibrosis Transmembrane Conductance Regulator misc Congenic Strain misc Cftr Gene misc Excision Event
topic_browse	misc Cystic Fibrosis misc Cystic Fibrosis Transmembrane Conductance Regulator misc Congenic Strain misc Cftr Gene misc Excision Event
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title	Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model
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title_full	Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model
author_sort	Charizopoulou, Nikoletta
journal	BMC genetics
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author_browse	Charizopoulou, Nikoletta Jansen, Silke Dorsch, Martina Stanke, Frauke Dorin, Julia R Hedrich, Hans-Jürgen Tümmler, Burkhard
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author-letter	Charizopoulou, Nikoletta
doi_str_mv	10.1186/1471-2156-5-6
title_sort	instability of the insertional mutation in cftrtgh(neoim)hgucystic fibrosis mouse model
title_auth	Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model
abstract	Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
abstractGer	Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
abstract_unstemmed	Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)HguCF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgumouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hguat the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. © Charizopoulou et al; licensee BioMed Central Ltd. 2004. This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
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title_short	Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model
url	https://dx.doi.org/10.1186/1471-2156-5-6
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author2	Jansen, Silke Dorsch, Martina Stanke, Frauke Dorin, Julia R Hedrich, Hans-Jürgen Tümmler, Burkhard
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Instability of the insertional mutation in CftrTgH(neoim)Hgucystic fibrosis mouse model

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