Please use this identifier to cite or link to this item: https://scholarbank.nus.edu.sg/handle/10635/33317
DC FieldValue
dc.titleA Baculovirus-Cre/1oxP Hybrid System for AAVS1 Locus-Directed Transgene Delivery
dc.contributor.authorCHRISHAN JULIAN ALLES RAMACHANDRA
dc.date.accessioned2012-05-31T18:01:35Z
dc.date.available2012-05-31T18:01:35Z
dc.date.issued2011-11-02
dc.identifier.citationCHRISHAN JULIAN ALLES RAMACHANDRA (2011-11-02). A Baculovirus-Cre/1oxP Hybrid System for AAVS1 Locus-Directed Transgene Delivery. ScholarBank@NUS Repository.
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/33317
dc.description.abstractInsertion of a transgene into a defined genomic locus in human embryonic stem cells (hESCs) is crucial in preventing random integration-induced insertional mutagenesis and can possibly enable persistent transgene expression during hESC expansion and in their differentiated progenies. Homologous recombination is first employed in hESCs to introduce heterospecific loxP sites into the AAVS1 locus, a site with an open chromatin structure that averts transgene silencing phenomena. Cre recombinase mediated cassette exchange is then performed using baculoviral vectors to insert a transgene into the modified AAVS1 locus. Targeting efficiency in the master hESC line with the loxP-docking sites was up to 100%. Expression of the inserted transgene lasted for at least 20 passages during hESC expansion and was retained in the differentiated progenies. Thus, this study demonstrates the feasibility of genome manipulation at the AAVS1 locus through the use of a baculovirus-Cre/loxP hybrid system. The technique developed will be useful for repeated gene targeting at a defined locus of the hESC genome.
dc.language.isoen
dc.subjectHuman embryonic stem cells, AAVS1 locus, Gene targeting, Homologous recombination, Recombinase-mediated cassette exchange, Baculoviral gene delivery
dc.typeThesis
dc.contributor.departmentBIOLOGICAL SCIENCES
dc.contributor.supervisorWANG SHU
dc.description.degreePh.D
dc.description.degreeconferredDOCTOR OF PHILOSOPHY
dc.identifier.isiutNOT_IN_WOS
Appears in Collections:Ph.D Theses (Open)

Show simple item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
RamachandraCJA.pdf1.51 MBAdobe PDF

OPEN

NoneView/Download

Page view(s)

511
checked on Sep 22, 2020

Download(s)

413
checked on Sep 22, 2020

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.