Let us imagine that some laboratory developed a method for creating restriction endonucleases that can specifically bind to short DNA sequence and specifically cut it or other determined DNA motif. In what applications would such enzymes be required? Could they be applied to Next Generation Sequencing?
If you use restriction endonucleases, describe their usage in few words or just say “I use them”.
I am aware of Zinc-Finger Nucleases or TALENs, but these enzymes only specifically bind target DNA and have non-specific DNA cutting activity.
Google "Zinc-Finger Nuclease" or "Transcription Activator-Like Effector Nuclease" (TALENs). You can buy these commercially (or design them yourself) to cut any DNA sequence you like.
Reality is more amazing than imagination any day.
Generally restriction endonuclease is used in day to day life in cloning purpose.
For cutting the the particular sequence of DNA and addition of desiracble fragment in that DNA,can be useful.
>a method for creation of restriction endonucleases specific to any desirable short DNA sequence.
I was already done and called - as Jason wrote - Zinc-Finger Nuclease.
http://en.wikipedia.org/wiki/Zinc_finger_nuclease
TALENs have 2 major disadvantages:
1.Domain responsible for G recognition also binds to A. Thus these enzymes cannot specifically recognise G-rich sequences.
2.Zinc-Finger Nucleases to some extent specifically bind to DNA target sequence, but they cut non-specifically around the target sequence.
These disadvantages lead to relatively high off-target activity of TALENs. And of course, next generation of endonuclease enzymes that can specifically recognize and specifically cut the target DNA are still required.
Its not just an idea. Engineering new restriction enzymes by directed evolution and modeling is being performed by numerous companies (such as NEB).
The raw idea is, indeed, use directed evolution combined with modeling. But I feel it might be the too long way to the result.
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