Stable Isogenic Recombinant (Sir) cell line technology
Generation of expression constructs
The Sir cell line system was developed for the stable single integration of DNA-constructs in an identical (isogenic) cellular context. By the use of special expression vectors we can integrate / express:
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Complete open reading frames
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Reporter constructs for cellular signaling pathways
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miRNAs and shRNA
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mutation constructs
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promoter regions
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TF-bindings sites
Requirements: The constructs to be expressed should be provided as sequence verified gateway entry clone.
For more information please contact: Dr. Rainer Will
Generation of acceptor cell lines
The isogenic cell line system is mainly based on the site specific recombination technology.
In the first instance a single recombination site is stably integrated in the nuclear genome of an immortalized mammalian cell line to create an “acceptor†cell line for site specific recombination. All available acceptor were tested for single integration and transcriptional activity.
Generation of expression cell lines
For the production of Sir expression cell lines genetic elements of interest are inserted into a single pre-defined genomic locus. This allows for the generation genetic twin cell lines sharing the identical genetic background except for the individual ‘ectopic’ genetic element that shall be expressed.
By site-specific recombination of ORFs (open reading frames), miRNAs, shRNAs or reporter gene constructs can be either inducible (CMV / Tet-on) or constitutively (EF1 or CMV promoter) expressed in the same genomic locus generating a highly standardized cellular environment.
