Document Detail


The chicken hypersensitivity site 4 core insulator blocks promoter interference in lentiviral vectors.
MedLine Citation:
PMID:  23448496     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Lentiviral vectors, including double internal promoters, can be used to express two transgenes in a single vector construct; however, transcriptional activities from double internal promoters are often inhibited by promoter interference. To determine whether the chicken hypersensitivity site 4 insulator (cHS4) could block promoter interference, lentiviral vectors including an MSCV-U3 promoter (Mp) and an EF1α promoter (Ep) were generated, and transgene expression was evaluated among transduced cells. In the Ep-Mp configuration, transcriptional activity from Mp was much lower, while Mp-Ep had similar transcription levels from both promoters. The cHS4 core insulator increased expression levels from Mp in HeLa cells, hematopoietic cell lines, and mouse peripheral blood cells following hematopoietic stem cell transplantation transduced with the Mp-Ep configured vector. This blocking function was mainly mediated by barrier activity regions in the insulator but not by CCCTC-binding factor (CTCF) binding sites. Cytosine-phosphate-guanine (CpG) methylation did not contribute to this barrier activity. In summary, combining the cHS4 insulator in double promoter vectors can improve transgene expression levels in various cell lines and mouse hematopoietic repopulating cells. These findings are useful for developing hematopoietic stem cell gene therapy.
Authors:
Naoya Uchida; Hideki Hanawa; Motoko Yamamoto; Takashi Shimada
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Intramural    
Journal Detail:
Title:  Human gene therapy methods     Volume:  24     ISSN:  1946-6544     ISO Abbreviation:  Hum Gene Ther Methods     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-04-11     Completed Date:  2013-10-17     Revised Date:  2014-04-01    
Medline Journal Info:
Nlm Unique ID:  101573202     Medline TA:  Hum Gene Ther Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  117-24     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Blood Cells / metabolism
Bone Marrow Cells / metabolism,  virology
Chickens / genetics
CpG Islands
DNA Methylation
Gene Expression
Gene Expression Regulation*
Gene Order
Genetic Vectors / genetics*
Humans
Insulator Elements*
Lentivirus / genetics*
Mice
Promoter Regions, Genetic*
RNA, Messenger / genetics
Transduction, Genetic
Transgenes
Chemical
Reg. No./Substance:
0/RNA, Messenger
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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