lenti-cre-nls

Lenti-ONE Cre-NLS

From €250.00

Description

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.

Specific integration of a sequence of interest into a LoxP site

Specific integration via Cre-NLS

The sequence of interest and the Cre recombinase are delivered by two non-integrative Lenti-ONE vectors.

Specific excision of a sequence flanked by LoxP sites

Specific excision via Cre-NLS

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.

Results

EXPRESSION OF CRE RECOMBINASES BY LENTI-ONE TRANS VECTORS IN CV1-5B CELLS MEDIATES EXCISION OF A TARGET SEQUENCE

In vitro application

LoxP site recombination in CV1-5B

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.

Transgenesis application

LoxP site recombination in Rosa26

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.