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.
Examples of Applications
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.
- Use in vitro, in vivo or for transgenesis purposes to mediate site specific integration of a substrate DNA vector which contains a LoxP site into a target LoxP site
- Use in vitro, in vivo or for transgenesis purposes to mediate excision of a DNA sequence flanked by LoxP sites, eg: to generate a conditional KO or to induce expression of a specific transgene
- Use in vitro, in vivo or for transgenesis purposes for Recombinase-Mediated Cassette Exchange in cells containing a target sequence flanked by LoxP sites
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.