Hi!
That looks good, the insert you painted has the neccessary recognition sequence so it will attach to the target and insert.
Basically it would be blunt ended then, right? What if you design your insert like ... No it is not blunt ended because your top insert N base-pairs with the bottom insert N.
The cell would surely recognize this as a strand break, and thus DNA ligase would ligate it. So I guess, as long as DNA ligase comes quicker than an exonuclease, the bacterium will survive. Most probably in the major part of the cells...
On Sunday, March 24, 2013 5:14:50 AM UTC+1, Koeng wrote:
Hello all!
I was just checking out some stuff about genome editing, but I couldn't really find anything about gene deletion inside a genome. Perhaps using a TALEN, would work is what I was thinking
A system goes like this: (the site it attaches to is also in the NNNNNNNN part)
ATATATAT'NNNNNNNN'GCGCGCGC
TATATATA'NNNNNNNN'CGCGCGCG
Then it is cut
ATAT CGCGGCGCGC
TATATATA CGCGCG
And I add some DNA
ATAT'N
N'GCGC
So it ligates
ATATATAT 'N' CGCGCGCG
TATATATA 'N' GCGCGCGC
Or what would the Bacteria do if its genome got cut like this? Die? Any documents for me to read would be helpful, or just your opinion. Thanks!
Koeng
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