Thank you so much. I knew nothing about Cre/LoxP, but now know something. It seems as complex, or more so than CRISPR. What purpose does Cre/LoxP serve in bacteriophage? Since viruses are so tiny, it's amazing that scientists were able to discover and study Cre/LoxP in phages.
If you are using CRISPR-Cas9, introduce the guide RNAs and a donor DNA template containing the loxP sites into the cells or organism of interest. The Cas9 enzyme will cut the DNA at the target sites, and the donor DNA will be integrated to repair the break and introduce the loxP sites. If you are using homologous recombination, introduce a DNA construct containing the loxP sites flanked by homology arms that match the upstream and downstream regions of the gene. The cells or organism will use the homology arms to swap the modified DNA with the endogenous DNA, introducing the loxP sites.
how do you turn the loxp site from right pointing to left pointing? I read the only thing you need to change is the 8bp core sequence but it was never clear to me how exactly one can change the sequence to invert the orientation
the best explanation I've had so far. Thank you!!!
You are most welcome ❤️
Thank you so much. I knew nothing about Cre/LoxP, but now know something. It seems as complex, or more so than CRISPR. What purpose does Cre/LoxP serve in bacteriophage? Since viruses are so tiny, it's amazing that scientists were able to discover and study Cre/LoxP in phages.
Best video on UA-cam for cre lox system
thanks
Great explanation. You are the best. Thank you
You are welcome.
thankyou this was very helpful mr!
A very helpful lecture , thanks a lot .
You are most welcome
Excellent lecture. You deserve a like
Thanks a lot!
It was very useful, thank you for your excellent teaching
You are most welcome 🤗
Thanks for the explanation. I have a question: how can I put the loxP sites upstream and downstream of a target gene?
If you are using CRISPR-Cas9, introduce the guide RNAs and a donor DNA template containing the loxP sites into the cells or organism of interest. The Cas9 enzyme will cut the DNA at the target sites, and the donor DNA will be integrated to repair the break and introduce the loxP sites.
If you are using homologous recombination, introduce a DNA construct containing the loxP sites flanked by homology arms that match the upstream and downstream regions of the gene. The cells or organism will use the homology arms to swap the modified DNA with the endogenous DNA, introducing the loxP sites.
@@BiologyLectures Thank you.
@@jinwon8471 you are most welcome.
thank you so much sir! 🤍
Very good lecture
Thanks for liking
how do you turn the loxp site from right pointing to left pointing? I read the only thing you need to change is the 8bp core sequence but it was never clear to me how exactly one can change the sequence to invert the orientation
did you find that please let me know
Excellent explanation sir 😊
Thanks and welcome