Transcriptional behavior of LCR enhancer elements integrated at the same chromosomal locus by recombinase-mediated cassette exchange

Eric E. Bouhassira, Karen Westerman, Philippe Leboulch

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

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 β-globin gene Locus Control Region by integrating at the same genetic locus in MEL cells, a LacZ gene driven by the human β-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 β- 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.

Original languageEnglish (US)
Pages (from-to)3332-3344
Number of pages13
JournalBlood
Volume90
Issue number9
StatePublished - 1997

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Recombinases
Globins
Genetic Loci
Transcription
Bearings (structural)
Genes
Locus Control Region
Cells
Hemin
Lac Operon
Aptitude
Chromosomes
Dimethyl Sulfoxide
Transgenes
Population

ASJC Scopus subject areas

  • Hematology

Cite this

Transcriptional behavior of LCR enhancer elements integrated at the same chromosomal locus by recombinase-mediated cassette exchange. / Bouhassira, Eric E.; Westerman, Karen; Leboulch, Philippe.

In: Blood, Vol. 90, No. 9, 1997, p. 3332-3344.

Research output: Contribution to journalArticle

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