Selection of target sites for antisense attack of RNA
View Patent ↗Nucleic acid hybridization under steady-state conditions is described by a kinetic model in which the intermediate state is assumed to be locally single stranded. An expression was derived that relates nucleic acid secondary structure to the rate of oligonucleotide-RNA hybridization. The model allows the calculation of a rate factor that is proportional to the rate constant for hybridization between complementary nucleic acids and is generally applicable to any RNA molecule with potential utility for rapid identification of sites for antisense attack of mRNA.
1. A method for identifying at least one target site of an RNA molecule for attack with an antisense RNA nucleic acid, the method comprising:
(a) for all short pairing regions in an RNA molecule, said short pairing regions having a selected length in the range of 11 to 20 nucleotides
(i) calculating the melting energy (ΔGm) required to convert each of said short pairing regions of the RNA molecule to a single-stranded state;
(ii) calculating the energy gain (ΔG d ) resulting from hybridization of each of said short pairing regions to an antisense RNA nucleic acid; and
(iii) calculating rate factor x wherein
x
=
1
C
ⅇ
Δ
G
m
/
RT
+
ⅇ
Δ
G
d
/
RT
,
wherein C is a proportionality constant, R is the universal gas constant and T is temperature;
(b) identifying at least one region that has a maximal value of rate factor x; and
(c) displaying the identified at least one region that has a maximal value of rate factor.
2. A device for identifying target sites of RNA molecules for attack with antisense RNA nucleic acids, the device comprising a computer having an executable program for carrying out the process steps of the method of claim 1 and displaying the identified at least one region that has a maximal value of rate factor.