Rosa26 mice conditionally express Neo or LacZ. Injection of Lenti-ONE Trans vector Cre-NLS in ROSA26 zygote induces LoxP site recombination, excision of the Neo cassette and expression of LacZ.
Lenti-ONE GFP carries the transgene green fluorescent protein. Its expression can be assessed by fluorescent microscopy or by flow cytometry, thus giving valuable informations about the transduction efficiency of the vector.
Cre-NLS cDNA encodes a cyclization recombinase fused with a nuclear localization signal (sequence from Enterobacterio phage P1 genome, AB449974.1/AX343218.1, 1053 pb). This tyrosine recombinase recognizes LoxP sequences and mediates recombination between two DNA fragments. This reaction can be used to excise a DNA fragment flanked by 2 loxP sequences or to integrate a DNA fragment into a single loxP sequence. Addition of a nuclear localization signal increases the efficiency of the enzyme.
The sequence of interest and the Cre recombinase are delivered by two non-integrative Lenti-ONE vectors.
Cre recombinase is delivered by a non-integrative Lenti-ONE vector
For this lentiviral vector, we recommend choosing a non-integrative mutant (IN(D64V) for example). This will make the Cre expression transient so that the potential toxicity of the recombinase is not deleterious.
If you wish to order a lentiviral vector with loxP sites, and the transgene that you need to integrate, please contact us and we will gladly produce it for you.
EXPRESSION OF CRE RECOMBINASES BY LENTI-ONE TRANS VECTORS IN CV1-5B CELLS MEDIATES EXCISION OF A TARGET SEQUENCE
In vitro application
CV1-5B cells conditionally express Neo or LacZ. Transduction of CV1-5B cell line with Lenti-ONE Trans vector Cre-NLS induces LoxP site recombination, excision of the Neo cassette and expression of LacZ.
Rosa26 mice conditionally express Neo or LacZ. Injection of Lenti-ONE Trans vector Cre-NLS in ROSA26 zygote induces LoxP site recombination, excision of the Neo cassette and expression of LacZ.
