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Conference Paper: Cre-transgenic mouse line for conditional gene knockout in retinal rod bipolar cells

TitleCre-transgenic mouse line for conditional gene knockout in retinal rod bipolar cells
Authors
KeywordsBipolar cells
Retinal development
Gene transfer/gene therapy
Issue Date2005
PublisherAssociation for Research in Vision and Ophthalmology. The Journal's web site is located at http://www.iovs.org
Citation
The 2005 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), Fort Lauderdale, FL., 1-5 May 2005. In Investigative Ophthalmology & Visual Science, 2005, v. 46 n. 13, p. 3111 How to Cite?
AbstractPURPOSE: Some progressive retinal diseases are caused by degeneration of neurons in the rod–pathway. It remains unknown how rod bipolar cells contribute to retinal degenerative disease. This study was conducted to establish a transgenic mouse line expressing Cre recombinase specifically in retinal rod bipolar cells. METHODS: IRES–Cre–cDNA fragment was inserted into a 173–kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene using Red–mediated recombineering. Transgenic mice were generated with the modified BAC and identified by Southern blot analysis and PCR. The activity of Cre–recombinase was analyzed in progenies from Cre–transgenic mice crossed with ROSA26 or Z/EG reporter mice. RESULTS: The genotypes of BAC–Pcp2–IRES–Cre transgenic mice were confirmed by PCR with Cre–specific primers. X–gal staining showed that the activities of Cre–recombinase were present in cerebellar Purkinje cells and retinal inner nuclear layer, where rod bipolar cells are located. Immunofluorescent double–staining with anti–GFP antibody and anti–PKCα (specific marker for retinal rod bipolar cells) antibody revealed that Cre–recombinase activity localized exclusively to the rod bipolar cells in the retina. CONCLUSIONS: A transgenic mouse line BAC–Pcp2–IRES–Cre that expresses Cre–recombinase in retinal rod bipolar cells has been successfully established. It will be a useful new tool for investigating the effects of retinal rod bipolar cell–specific gene inactivation and invaluable for future studies on retinal functions.
DescriptionThis free journal issue entitled: ARVO Annual Meeting Abstract
Persistent Identifierhttp://hdl.handle.net/10722/95045
ISSN
2015 Impact Factor: 3.427
2015 SCImago Journal Rankings: 2.008

 

DC FieldValueLanguage
dc.contributor.authorHuang, J-
dc.contributor.authorZhang, X-
dc.contributor.authorChen, B-
dc.contributor.authorNg, AHL-
dc.contributor.authorTanner, JA-
dc.contributor.authorTay, DKC-
dc.contributor.authorSo, KF-
dc.contributor.authorRachel, RA-
dc.contributor.authorCopeland, NG-
dc.contributor.authorJenkins, NA-
dc.date.accessioned2010-09-25T15:49:55Z-
dc.date.available2010-09-25T15:49:55Z-
dc.date.issued2005-
dc.identifier.citationThe 2005 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), Fort Lauderdale, FL., 1-5 May 2005. In Investigative Ophthalmology & Visual Science, 2005, v. 46 n. 13, p. 3111-
dc.identifier.issn0146-0404-
dc.identifier.urihttp://hdl.handle.net/10722/95045-
dc.descriptionThis free journal issue entitled: ARVO Annual Meeting Abstract-
dc.description.abstractPURPOSE: Some progressive retinal diseases are caused by degeneration of neurons in the rod–pathway. It remains unknown how rod bipolar cells contribute to retinal degenerative disease. This study was conducted to establish a transgenic mouse line expressing Cre recombinase specifically in retinal rod bipolar cells. METHODS: IRES–Cre–cDNA fragment was inserted into a 173–kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene using Red–mediated recombineering. Transgenic mice were generated with the modified BAC and identified by Southern blot analysis and PCR. The activity of Cre–recombinase was analyzed in progenies from Cre–transgenic mice crossed with ROSA26 or Z/EG reporter mice. RESULTS: The genotypes of BAC–Pcp2–IRES–Cre transgenic mice were confirmed by PCR with Cre–specific primers. X–gal staining showed that the activities of Cre–recombinase were present in cerebellar Purkinje cells and retinal inner nuclear layer, where rod bipolar cells are located. Immunofluorescent double–staining with anti–GFP antibody and anti–PKCα (specific marker for retinal rod bipolar cells) antibody revealed that Cre–recombinase activity localized exclusively to the rod bipolar cells in the retina. CONCLUSIONS: A transgenic mouse line BAC–Pcp2–IRES–Cre that expresses Cre–recombinase in retinal rod bipolar cells has been successfully established. It will be a useful new tool for investigating the effects of retinal rod bipolar cell–specific gene inactivation and invaluable for future studies on retinal functions.-
dc.languageeng-
dc.publisherAssociation for Research in Vision and Ophthalmology. The Journal's web site is located at http://www.iovs.org-
dc.relation.ispartofInvestigative Ophthalmology & Visual Science-
dc.subjectBipolar cells-
dc.subjectRetinal development-
dc.subjectGene transfer/gene therapy-
dc.titleCre-transgenic mouse line for conditional gene knockout in retinal rod bipolar cells-
dc.typeConference_Paper-
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0146-0404&volume=46&spage=3111S&epage=&date=2005&atitle=Cre-transgenic+mouse+line+for+conditional+gene+knockout+in+retinal+rod+bipolar+cells+en_HK
dc.identifier.emailHuang, J: jdhuang@hkucc.hku.hk-
dc.identifier.emailZhang, X: zhangxm@hkucc.hku.hk-
dc.identifier.emailTanner, JA: jatanner@hkucc.hku.hk-
dc.identifier.emailTay, DKC: dkctay@hkucc.hku.hk-
dc.identifier.emailSo, KF: hrmaskf@hkucc.hku.hk-
dc.identifier.authorityHuang, J=rp00451-
dc.identifier.authorityTanner, JA=rp00495-
dc.identifier.authorityTay, DKC=rp00336-
dc.identifier.authoritySo, KF=rp00329-
dc.identifier.hkuros100385-
dc.identifier.volume46-
dc.identifier.issue13-
dc.identifier.spage3111-
dc.identifier.epage3111-
dc.publisher.placeUnited States-
dc.customcontrol.immutablesml 170407 amended-

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