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The Hairpin Ribozyme: Discovery and Development for Gene Therapy

 Last Update: 2021-01-31
 Source: Internet
 Author: User

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The minimum catalytic center of (-)s TRSV was identified, biochemically characterized and named the hairpin ribozyme (
1
,
2
). Following initial identification of the minimum catalytic sequence, we identified a
trans
-catalytic reaction and biochemically characterized this reaction. The minimum sequence for catalytic activity was found to be a 50 nucleotide (nt) ribozyme and a 14 nt substrate. This was determined by modeling and cut-down experiments. A 50 nt catalytic RNA could cleave a 14 nt substrate RNA and the reaction occurred in
trans.
The reaction proceeded without depletion of the 50 nt RNA component, and therefore was catalytic. It had true Michaelis-Menten kinetics allowing determination of K
M
,k
cat
, energy of activation, Mg
2+
dependence, and the pH optima.

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