|
|
 Previous Article | Table of Contents | Next Article 
RAPID COMMUNICATION
Transcriptional Behavior of LCR Enhancer Elements Integrated at the Same Chromosomal Locus by Recombinase-Mediated Cassette Exchange
Eric E. Bouhassira,
Karen Westerman, and
Philippe Leboulch
From the Division of Hematology, the Department of Medicine, Albert Einstein College of Medicine, Bronx, NY; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA; and Harvard Medical School, the Department of Medicine, Brigham & Women's Hospital, Boston, MA.
Efficient integration of transgenes at preselected chromosomal locations was achieved in mammalian cells by recombinase-mediated-cassette-exchange (RMCE), a novel procedure that makes use of the CRE recombinase together with Lox sites bearing different spacer regions. We have applied RMCE to the study of the human  -globin gene Locus Control Region by integrating at the same genetic locus in MEL cells, a LacZ gene driven by the human -globin promoter linked to HS2 and HS3 alone or in combination with HS4. Expression studies at the cell population level and in individual cells before and after induction of differentiation with hemin or DMSO show that the presence of these enhancers is associated with variegated patterns of expression. We were able to show that the LCR fragments tested act by controlling both the probability of expression and the rate of transcription of the linked -globin promoter. Both of these factors were also dependent on the state of differentiation of the MELc and on the presence of a second transcription unit located in cis. The ability to manipulate by RMCE constructs integrated into chromosomes should help in the creation of complex, rationally designed, artificial genetic loci.
Blood, Vol. 90 No. 9 (November 1), 1997:
pp. 3332-3344
© 1997 by The American Society of Hematology.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
J. D. Louwerse, M. C.M. van Lier, D. M. van der Steen, C. M.T. de Vlaam, P. J.J. Hooykaas, and A. C. Vergunst
Stable Recombinase-Mediated Cassette Exchange in Arabidopsis Using Agrobacterium tumefaciens
Plant Physiology,
December 1, 2007;
145(4):
1282 - 1293.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. J. T. Venken and H. J. Bellen
Transgenesis upgrades for Drosophila melanogaster
Development,
October 15, 2007;
134(20):
3571 - 3584.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Ghosh, F. Guo, and G. D. Van Duyne
Synapsis of loxP Sites by Cre Recombinase
J. Biol. Chem.,
August 17, 2007;
282(33):
24004 - 24016.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Wang, Y. Zhang, Y. Cheng, Y. Zhou, D. C. King, J. Taylor, F. Chiaromonte, J. Kasturi, H. Petrykowska, B. Gibb, et al.
Experimental validation of predicted mammalian erythroid cis-regulatory modules
Genome Res.,
December 1, 2006;
16(12):
1480 - 1492.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Araki, M. Araki, and K.-i. Yamamura
Negative Selection with the Diphtheria toxin A fragment Gene Improves Frequency of Cre-Mediated Cassette Exchange in ES Cells
J. Biochem.,
December 1, 2006;
140(6):
793 - 798.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Toledo, C.-W. Liu, C. J. Lee, and G. M. Wahl
RMCE-ASAP: a gene targeting method for ES and somatic cells to accelerate phenotype analyses
Nucleic Acids Res.,
July 26, 2006;
34(13):
e92 - e92.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. R. Bateman, A. M. Lee, and C.-t. Wu
Site-Specific Transformation of Drosophila via {phi}C31 Integrase-Mediated Cassette Exchange
Genetics,
June 1, 2006;
173(2):
769 - 777.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. T. Wong, J. L Kolman, Y.-C. Li, L. D Mesner, W. Hillen, C. Berens, and G. M Wahl
Reproducible doxycycline-inducible transgene expression at specific loci generated by Cre-recombinase mediated cassette exchange
Nucleic Acids Res.,
October 4, 2005;
33(17):
e147 - e147.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Nakano, K. Odaka, Y. Takahashi, M. Ishimura, I. Saito, and Y. Kanegae
Production of viral vectors using recombinase-mediated cassette exchange
Nucleic Acids Res.,
May 5, 2005;
33(8):
e76 - e76.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Masui, D. Shimosato, Y. Toyooka, R. Yagi, K. Takahashi, and H. Niwa
An efficient system to establish multiple embryonic stem cell lines carrying an inducible expression unit
Nucleic Acids Res.,
March 1, 2005;
33(4):
e43 - e43.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. B. Osipovich, A. Singh, and H. E. Ruley
Post-entrapment genome engineering: First exon size does not affect the expression of fusion transcripts generated by gene entrapment
Genome Res.,
March 1, 2005;
15(3):
428 - 435.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S.-H. L. Kang, P. P Levings, F. Andersen, P. J Laipis, K. I Berns, R. T Zori, and J. Bungert
Locus control region elements HS2 and HS3 in combination with chromatin boundaries confer high-level expression of a human {beta}-globin transgene in a centromeric region
Genes Cells,
November 1, 2004;
9(11):
1043 - 1053.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. A. Adelman, S. Chattopadhyay, and J. J. Bieker
The BMP/BMPR/Smad pathway directs expression of the erythroid-specific EKLF and GATA1 transcription factors during embryoid body differentiation in serum-free media
Development,
March 3, 2003;
129(2):
539 - 549.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Imren, E. Payen, K. A. Westerman, R. Pawliuk, M. E. Fabry, C. J. Eaves, B. Cavilla, L. D. Wadsworth, Y. Beuzard, E. E. Bouhassira, et al.
Permanent and panerythroid correction of murine beta thalassemia by multiple lentiviral integration in hematopoietic stem cells
PNAS,
October 29, 2002;
99(22):
14380 - 14385.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Araki, M. Araki, and K.-i. Yamamura
Site-directed integration of the cre gene mediated by Cre recombinase using a combination of mutant lox sites
Nucleic Acids Res.,
October 1, 2002;
30(19):
e103 - e103.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. J. Langer, A. P. Ghafoori, M. Byrd, and L. Leinwand
A genetic screen identifies novel non-compatible loxP sites
Nucleic Acids Res.,
July 15, 2002;
30(14):
3067 - 3077.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Pawliuk, K. A. Westerman, M. E. Fabry, E. Payen, R. Tighe, E. E. Bouhassira, S. A. Acharya, J. Ellis, I. M. London, C. J. Eaves, et al.
Correction of Sickle Cell Disease in Transgenic Mouse Models by Gene Therapy
Science,
December 14, 2001;
294(5550):
2368 - 2371.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. M. Molete, H. Petrykowska, E. E. Bouhassira, Y.-Q. Feng, W. Miller, and R. C. Hardison
Sequences Flanking Hypersensitive Sites of the {beta}-Globin Locus Control Region Are Required for Synergistic Enhancement
Mol. Cell. Biol.,
May 1, 2001;
21(9):
2969 - 2980.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
A. Ramírez, E. Milot, I. Ponsa, C. Marcos-Gutiérrez, A. Page, M. Santos, J. Jorcano, and M. Vidal
Sequence and Chromosomal Context Effects on Variegated Expression of Keratin 5/lacZ Constructs in Stratified Epithelia of Transgenic Mice
Genetics,
May 1, 2001;
158(1):
341 - 350.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
Y.-Q. Feng, M. C. Lorincz, S. Fiering, J. M. Greally, and E. E. Bouhassira
Position Effects Are Influenced by the Orientation of a Transgene with Respect to Flanking Chromatin
Mol. Cell. Biol.,
January 1, 2001;
21(1):
298 - 309.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
E. C. Forsberg, T. N. Zaboikina, W. K. Versaw, N. G. Ahn, and E. H. Bresnick
Enhancement of beta -Globin Locus Control Region-Mediated Transactivation by Mitogen-Activated Protein Kinases through Stochastic and Graded Mechanisms
Mol. Cell. Biol.,
August 1, 1999;
19(8):
5565 - 5575.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y.-Q. Feng, R. Alami, and E. E. Bouhassira
Enhancer-Dependent Transcriptional Oscillations in Mouse Erythroleukemia Cells
Mol. Cell. Biol.,
July 1, 1999;
19(7):
4907 - 4917.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. C. Walters, S. Fiering, E. E. Bouhassira, D. Scalzo, S. Goeke, W. Magis, D. Garrick, E. Whitelaw, and D. I. K. Martin
The Chicken beta -Globin 5'HS4 Boundary Element Blocks Enhancer-Mediated Suppression of Silencing
Mol. Cell. Biol.,
May 1, 1999;
19(5):
3714 - 3726.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Esperet, S. Sabatier, M.-A. Deville, R. Ouazana, E. E. Bouhassira, J. Godet, F. Morle, and A. Bernet
Non-erythroid Genes Inserted on Either Side of Human HS-40 Impair the Activation of Its Natural alpha -Globin Gene Targets without Being Themselves Preferentially Activated
J. Biol. Chem.,
August 11, 2000;
275(33):
25831 - 25839.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Elnitski, J. Li, C. T. Noguchi, W. Miller, and R. Hardison
A Negative Cis-element Regulates the Level of Enhancement by Hypersensitive Site 2 of the beta -Globin Locus Control Region
J. Biol. Chem.,
February 23, 2001;
276(9):
6289 - 6298.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
