The Cre-lox and FLP-FRT systems. Two of the most exciting and versatile genetic tools designed in the last 30 years are the Cre-lox and FLP-FRT technologies. Both allow the location and timing of gene expression to be closely regulated. This article briefly outlines how the two systems work and can be used in mouse models. Flp-In System: For Generating Constitutive Expression Cell Lines. For Generating Constitutive Expression Cell Lines be sure to use supercoiled pOG44 and pcDNA5/FRT plasmid DNA. Flp-mediated recombination between the FRT site on pcDNA5/FRT and the integrated FRT site in the Flp-In™ host cell line will only occur if the pcDNA5/FRT. The Flp-In™ System allows for stable integration and expression of your gene of interest to deliver single-copy isogenic cell lines. Flp-In™ expression involves introduction of a Flp Recombination Target (FRT) site into the genome of the mammalian cell line of choice. Most mouse geneticists who work with Cre-lox mice probably are familiar with the FLP-FRT system. Like Cre, FLP is a tyrosine recombinase, but it originates from the yeast Saccharomyces cerevisiae, rather than from bacteriophage. In FLP-FRT, FLP is analogous to cre and FRT to loxP – FLP recognizes the FRT sites, and catalyzes recombination. Jun 15, · To assay the efficiency of the FLP/FRT site-specific recombination system in Danio rerio, FLPe capped RNA was microinjected into transgenic zebrafish embryos carrying an FRT-flanked EGFP expression cassette. Injected fish were observed for muscle-specific GFP expression, and outcrossed to wild-type zebrafish to assess germline excision yourabout.com by: In zebrafish, transgenesis-based recombinase genetics using Cre/lox, Flp/FRT, and ΦC31 are increasingly applied to study development and homeostasis, and to generate disease models. FLP Project Free. 88 likes. Free Remakes FLP Project Free For request contact: [email protected] -Remake -Template -Tutorial -Ghost ProductionFollowers: Aug 01, · Two publications in reported FLP/FRT-based excision systems in C. elegans: Davis et al., () 15 and Voutev and Hubbard () Both demonstrate spatial and temporal control of somatic gene expression, using either heat-shock inducible or spatially-restricted recombinase expression combined with ubiquitous or spatially-restricted Cited by:
These arrays contained a positive selection marker flanked by loxP sites. Received Mar 17; Accepted May 8. For example, additional layer of control could be imposed by regulation of heat-shock response by restricted expression of hsf-1 Reverse-transcription PCR and northern blotting confirmed the excision of the 18th exon of tm1Cwr abundantly in the brain tissue and moderately in muscular tissue due to Scwhann cells within the muscle. Efficient expression of codon-adapted genes in filamentous fungi.
Van Duyne, G. Andreeva, C. PCR analysis of S. Drosophila Brainbow: a recombinase-based fluorescence labeling technique to subdivide neural expression patterns. This article has been cited by other articles in PMC.
Further studies will determine the basis of these effects and hopefully provide better guidelines for selecting promoters and recombinases that are optimal for regulating gene expression in a particular cell of interest. Espagne, K. Gene activation using FLP recombinase in C. Gustafsson, C. The Caenorhabditis elegans APC-related gene apr-1 is required for epithelial cell migration and Hox gene expression.