What is FLEx Cre?
What Are FLEx switches? FLEx (or “flip-excision”) switches were designed as a genetic tool for researchers to conditionally manipulate gene expression in vivo using site-specific recombination. The FLEx switch takes advantage of the orientation specificity of the site-specific recombinases (SSRs) Cre and FLP.
What does Cre loxP do?
The Cre-lox system is used as a genetic tool to control site specific recombination events in genomic DNA. This system has allowed researchers to manipulate a variety of genetically modified organisms to control gene expression, delete undesired DNA sequences and modify chromosome architecture.
How does the Cre loxP system work?
How does it work? As alluded to above, the Cre recombinase catalyzes the site specific recombination event between two loxP sites, which can be located either on the same or on separate pieces of DNA. Both 13bp repeat sequences on a single loxP site are recognized and bound by a Cre protein, forming a dimer.
Is FLEx Cre dependent?
We have developed a widely used Cre-dependent viral vector utilizing a genetic element called a FLEX switch. These vectors are prone to recombination. Therefore, to minimize recombination, we propagate these plasmids in Stbl2 cells from Invitrogen.
What is Dio in virus?
The double-floxed inverse open reading frame (DIO) construct comprises inverted terminal repeats (ITR), the EF1α promoter, an eYFP-ChR2 fusion gene surrounded by a pair of LoxP sites and a pair of Lox2722 sites oriented inward, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and a human …
What is plasmid Wpre?
Woodchuck Hepatitis Virus (WHV) Posttranscriptional Regulatory Element (WPRE) is a DNA sequence that, when transcribed, creates a tertiary structure enhancing expression. The sequence is commonly used in molecular biology to increase expression of genes delivered by viral vectors.
Is Cre lox reversible?
Cre/loxP-mediated inversion of genomic DNA segments represents a useful tool to alter the mouse genome in a cell type- and/or tissue-specific manner. However, its applicability is hampered by the fact that this reaction is fully reversible.
Can be studied using Cre loxP system?
In particular, Cre-loxP system is widely used as an integral experimental tool for generating the conditional. This system has enabled researchers to investigate genes of interest in a tissue/cell (spatial control) and/or time (temporal control) specific manner.
What does double Floxed mean?
In genetics, floxing refers to the sandwiching of a DNA sequence (which is then said to be floxed) between two lox P sites. Floxing a gene allows it to be deleted (knocked out), translocated or inverted in a process called Cre-Lox recombination.
Is WPRE necessary?
The WPRE is often essential to achieve sufficient levels of expression with integrative retroviral vectors configured for gene therapy; therefore, its mechanisms of action have been studied in detail.