YILIMART - StemMACS Cre Recombinase mRNA

StemMACS™ Cre Recombinase mRNA encodes a codon-optimized variant of Cre recombinase for transient transfection.

Applications

site-specific recombination between loxP sites

Background information

Cre recombinase is a tyrosine recombinase enzyme derived from the P1 bacteriophage that catalyzes site-specific recombination between two DNA recognition sites (loxP sites).

Detailed procedure

For satisfactory transfection results, use a transfection protocol optimized for your specific cell type. StemMACS™ eGFP mRNA or StemMACS™ Nuclear eGFP mRNA allow easy evaluation of transfection efficiency and are recommended as positive controls.

Quality description

StemMACS™ mRNAs are highly pure, in vitro-transcribed mRNAs that have been carefully optimized to ensure high level expression after transfection. Polyadenylation and enzymatic cap mimic a fully processed mature mRNA and ensure efficient translation. Incorporation of the modified nucleotides pseudouridine and 5-methylcytidine reduces the innate antiviral response to single-stranded mRNA. Robust expression of the encoded factor after transfection has been verified by immunofluorescence or flow cytometry.

StemMACS™ mRNAs are lyophilized from a 0.2 µM filtered solution for extended shelf-life.

Figure 1

Cre-mediated recombination in mouse ES cells:

HM1 embryonic stem cells carrying a GFP reporter gene that is transcribed upon cre-mediated recombination were electroporated with 3 μg StemMACS™ Cre Recombinase mRNA and analyzed by flow cytometry 24 hours after transfection. 72% of all cells were GFP-positive indicating successful cre-mediated recombination.

Untransfected cells	Transfected cells

Figure 2
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Figure 2

Full control over recombination efficiency:
The optimal amount of StemMACS™ Cre Recombinase mRNA can be determined by titration. In the case of the HM1 reporter line described above, a single electroporation with >3 μg StemMACS™ Cre Recombinase mRNA resulted in 70-80% recombination. Transfection efficiency was >80% as determined by independent electroporation of StemMACS™ eGFP mRNA into control HM1 cells (not shown).

Full control over recombination efficiency:
The optimal amount of StemMACS™ Cre Recombinase mRNA can be determined by titration. In the case of the HM1 reporter line described above, a single electroporation with >3 μg StemMACS™ Cre Recombinase mRNA resulted in 70-80% recombination. Transfection efficiency was >80% as determined by independent electroporation of StemMACS™ eGFP mRNA into control HM1 cells (not shown).

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StemMACS Cre Recombinase mRNA

For research use only

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