PARTS

Overview

In the previous section of DNAzyme mutation, we have mentioned that in pursuit of better performance, we mutated 8-17 DNAzyme and obtained a series of mutants. For these mutants, we conducted a lot of tests and finally screened out a DNAzyme with better performance than 8-17.

Design

Fig. 1 2D structure of 8-17 DNAzyme

For mutant 1-9, we replaced a single base pair on their large stem ring.
For mutant 10-24, we replaced a single base pair on their small stem ring.
For mutant 25-40, we inserted a single base pair into their large stem ring.
For mutant 41-64, we inserted a single base pair into their large stem ring.
For mutant 65-67, we reversed the base pairs on their small stem ring.
For mutant 68-70, we modified the base pairs on their small stem ring.
For mutant 71-79, we modified the base pairs on the end of their stem rings.
For mutant 80-83, we elongated the base pairs on their large stem ring.
For mutant 84-87, we elongated the base pairs on their small stem ring.
For mutant 88-95, we elongated the base pairs on their stem rings.

Result

We found that 14 of them have DNAzyme activity, and among these 14 types, mut 10 has similar effects to 8-17.

Fig. 2 8-17 DNAzyme mutation test

Selected 14 mutants:

mut 9, mut 10, mut 11, mut 12, mut 20, mut 21, mut 23, mut 24, mut 35, mut 39, mut 60, mut 64, mut 65, mut 70.

Fig. 3 Further testing of 14 candidate mutations.

Then we compared the detection limit of the original DNAzyme and mut 10. The results are followed

Fig. 4 DNAzyme dectection test. Left: mut 10; Right: Original

Conclusion:

Through mutation experiments, we screened out a large number of mutants with DNAzyme activity. And among these mutants, Mut 10 has better shear activity and metal ion sensitivity

Parts Collection: