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Evaluation of CRE-mediated excision approaches in Arabidopsis thaliana

Gordana Marjanac UGent, Annelies De Paepe UGent, Ingrid Peck UGent, Anni Jacobs UGent, Sylvie De Buck UGent and Anna Depicker UGent (2008) TRANSGENIC RESEARCH. 17(2). p.239-250
abstract
The ability of the CRE recombinase to catalyze excision of a DNA fragment flanked by directly repeated lox sites has been exploited to modify gene expression and proved to function well in particular case studies. However, very often variability in CRE expression and differences in efficiency of CRE-mediated recombination are observed. Here, various approaches were investigated to reproducibly obtain optimal CRE activity. CRE recombination was analyzed either by transforming the CRE T-DNA into plants containing a lox-flanked fragment or by transforming a T-DNA harboring a lox-flanked fragment into plants producing the CRE recombinase. Although somatic CRE-mediated excision of a lox-flanked fragment was obtained in all transformants, a variable amount of germline-transmitted deletions was found among different independent transformants, irrespective of the orientation of transformation. Also, the efficiency of CRE-mediated excision correlated well with the CRE mRNA level. In addition, CRE-mediated fragment excision was compared after floral dip and after root tissue transformation when transforming in a CRE-expressing background. Importantly, less CRE activity was needed to excise the lox-flanked fragment from the transferred T-DNA after root tissue transformation than after floral dip transformation. We hypothesize that this is correlated with the lower T-DNA copy number inserted during root transformation as compared to floral dip transformation.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
TRANSFORMATION, TRANSIENT EXPRESSION, TOBACCO, SYSTEM, ROOT EXPLANTS, TRANSGENIC PLANTS, SELECTABLE MARKER GENE, GLUCURONIDASE ACCUMULATION LEVELS, T-DNA INTEGRATION, SITE-SPECIFIC RECOMBINATION, T-DNA, root transformation, floral dip, Arabidopsis, CRE/loxP recombination, excision
journal title
TRANSGENIC RESEARCH
Transgenic Res.
volume
17
issue
2
pages
239 - 250
Web of Science type
Article
Web of Science id
000254353900008
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
2.809 (2008)
JCR rank
43/144 (2008)
JCR quartile
2 (2008)
ISSN
0962-8819
DOI
10.1007/s11248-007-9096-9
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
409425
handle
http://hdl.handle.net/1854/LU-409425
date created
2008-05-15 18:03:00
date last changed
2013-10-21 11:05:33
@article{409425,
  abstract     = {The ability of the CRE recombinase to catalyze excision of a DNA fragment flanked by directly repeated lox sites has been exploited to modify gene expression and proved to function well in particular case studies. However, very often variability in CRE expression and differences in efficiency of CRE-mediated recombination are observed. Here, various approaches were investigated to reproducibly obtain optimal CRE activity. CRE recombination was analyzed either by transforming the CRE T-DNA into plants containing a lox-flanked fragment or by transforming a T-DNA harboring a lox-flanked fragment into plants producing the CRE recombinase. Although somatic CRE-mediated excision of a lox-flanked fragment was obtained in all transformants, a variable amount of germline-transmitted deletions was found among different independent transformants, irrespective of the orientation of transformation. Also, the efficiency of CRE-mediated excision correlated well with the CRE mRNA level. In addition, CRE-mediated fragment excision was compared after floral dip and after root tissue transformation when transforming in a CRE-expressing background. Importantly, less CRE activity was needed to excise the lox-flanked fragment from the transferred T-DNA after root tissue transformation than after floral dip transformation. We hypothesize that this is correlated with the lower T-DNA copy number inserted during root transformation as compared to floral dip transformation.},
  author       = {Marjanac, Gordana and De Paepe, Annelies and Peck, Ingrid and Jacobs, Anni and De Buck, Sylvie and Depicker, Anna},
  issn         = {0962-8819},
  journal      = {TRANSGENIC RESEARCH},
  keyword      = {TRANSFORMATION,TRANSIENT EXPRESSION,TOBACCO,SYSTEM,ROOT EXPLANTS,TRANSGENIC PLANTS,SELECTABLE MARKER GENE,GLUCURONIDASE ACCUMULATION LEVELS,T-DNA INTEGRATION,SITE-SPECIFIC RECOMBINATION,T-DNA,root transformation,floral dip,Arabidopsis,CRE/loxP recombination,excision},
  language     = {eng},
  number       = {2},
  pages        = {239--250},
  title        = {Evaluation of CRE-mediated excision approaches in Arabidopsis thaliana},
  url          = {http://dx.doi.org/10.1007/s11248-007-9096-9},
  volume       = {17},
  year         = {2008},
}

Chicago
Marjanac, Gordana, Annelies De Paepe, Ingrid Peck, Anni Jacobs, Sylvie De Buck, and Anna Depicker. 2008. “Evaluation of CRE-mediated Excision Approaches in Arabidopsis Thaliana.” Transgenic Research 17 (2): 239–250.
APA
Marjanac, G., De Paepe, A., Peck, I., Jacobs, A., De Buck, S., & Depicker, A. (2008). Evaluation of CRE-mediated excision approaches in Arabidopsis thaliana. TRANSGENIC RESEARCH, 17(2), 239–250.
Vancouver
1.
Marjanac G, De Paepe A, Peck I, Jacobs A, De Buck S, Depicker A. Evaluation of CRE-mediated excision approaches in Arabidopsis thaliana. TRANSGENIC RESEARCH. 2008;17(2):239–50.
MLA
Marjanac, Gordana, Annelies De Paepe, Ingrid Peck, et al. “Evaluation of CRE-mediated Excision Approaches in Arabidopsis Thaliana.” TRANSGENIC RESEARCH 17.2 (2008): 239–250. Print.