How is it that these lox sites flank genes of interest without including other genes? It seems unlikely that so many genes are flanked by palindromic sequences, but am I incorrect in thinking that?
These lox sites are typically inserted into expression vectors and genomes by researchers in very precise locations. They do not exist naturally as flanking sequences of particular genes. Researchers will determine where they want the inversions to occur, and will either clone in the lox sites into their plasmids, or perform knock-in experiments to insert the lox sequences in the desired positions in the genome before carrying out their Cre-induced inversions. Hope this helps!
Cre é projetada para um tipo de célula em específico? É preciso encontrar um promotor específico de uma determinada célula que deeja a marcação e então produzir uma cre recombinante própria para ser ativada por esse promotr em específico?
Nicely explained!! I have a question..how to incorporate cre sequence in a mouse at the gene specific promoter? And also the lox p sites at the specific position?
Depending on whether you would like to overexpress the cre sequence using a gene specific promoter, or to knock-in the cre sequence under the endogenous promoter, gene overexpression constructs or CRISPR knock-in can be used. If you have a specific project in mind, please contact technical@abmgood.com for further discussion.
The gene must be inverted in the OFF (no expression) condition for the FLEx system. The Cre-Lox FLEx system controls expression by changing the orientation of the gene in relation to its intended promoter. For example, in the OFF position, the promoter would be in the forward direction while the gene (flanked by Lox sites) would be in the reverse direction, which would not lead to expression of the functional gene. After Cre is introduced to turn the system ON, the gene would be flipped into the forward direction and be able to be expressed under the control of the promoter. Hope this helps!
This animation is enlightening.
Cre-loxP system is indeed an intricate wonder.
Thank you SO MUCH. Why in my university there are not videos like this one?
GREAT JOB and what an incredible genomic tecnique!
Hi Matteo, thanks for watching and leaving such a nice comment! Please let us know if you have any questions and we'd be glad to help!
Very helpful! Greetings from Brazil.
We're glad to hear that!
Amazing video! Why is it called a FLEx switch if it's irreversible? You can't toggle it off after inversion, right?
Dr. Moreau approves this video
Very useful! Thank you so much!
Extremely helpful video, THANKS!
You're welcome. Thanks for watching!
it was so helpful thanks a lot
You are welcome! Glad it helped!
How is it that these lox sites flank genes of interest without including other genes? It seems unlikely that so many genes are flanked by palindromic sequences, but am I incorrect in thinking that?
These lox sites are typically inserted into expression vectors and genomes by researchers in very precise locations. They do not exist naturally as flanking sequences of particular genes. Researchers will determine where they want the inversions to occur, and will either clone in the lox sites into their plasmids, or perform knock-in experiments to insert the lox sequences in the desired positions in the genome before carrying out their Cre-induced inversions. Hope this helps!
@@abmgood This is super helpful, thank you very much!
Great video and explanation, really helpful!
Thanks for watching!
Cre é projetada para um tipo de célula em específico? É preciso encontrar um promotor específico de uma determinada célula que deeja a marcação e então produzir uma cre recombinante própria para ser ativada por esse promotr em específico?
such a helpful video! Thank you!
Hello Caroline,
Thanks for watching! We are really glad to hear that our video was helpful :)
Now I can continue my article.. thanks!
You're welcome ;) Thanks for watching!
Thank you
Nicely explained!! I have a question..how to incorporate cre sequence in a mouse at the gene specific promoter? And also the lox p sites at the specific position?
Depending on whether you would like to overexpress the cre sequence using a gene specific promoter, or to knock-in the cre sequence under the endogenous promoter, gene overexpression constructs or CRISPR knock-in can be used. If you have a specific project in mind, please contact technical@abmgood.com for further discussion.
Really short n sweet
tysm!!
This video was very helpful.
Hi Mayukh, thanks for watching our video! Please let us know if you have any questions and we'd be glad to help!
It helps to clear my basics
Hi Sumitra, thanks for watching and glad our video was helpful - please let us know if you have any questions and we'd be glad to help!
very informative!
Thanks for watching! :)
Thank you! :)
You're welcome!
amazing explication!
Thank you! :)
Why does it matter if a gene is inverted?
The gene must be inverted in the OFF (no expression) condition for the FLEx system.
The Cre-Lox FLEx system controls expression by changing the orientation of the gene in relation to its intended promoter. For example, in the OFF position, the promoter would be in the forward direction while the gene (flanked by Lox sites) would be in the reverse direction, which would not lead to expression of the functional gene. After Cre is introduced to turn the system ON, the gene would be flipped into the forward direction and be able to be expressed under the control of the promoter. Hope this helps!
I know I have the genes to be sexy. I just need the switches to be flipped
great joke xD
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