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Stability and mismatch discrimination of locked nucleic acid-DNA duplexes Owczarzy R; You Y; Groth CL; Tataurov AVBiochemistry 2011[Nov]; 50 (43): 9352-67Locked nucleic acids (LNA; symbols of bases, +A, +C, +G, and +T) are introduced into chemically synthesized oligonucleotides to increase duplex stability and specificity. To understand these effects, we have determined thermodynamic parameters of consecutive LNA nucleotides. We present guidelines for the design of LNA oligonucleotides and introduce free online software that predicts the stability of any LNA duplex oligomer. Thermodynamic analysis shows that the single strand-duplex transition is characterized by a favorable enthalpic change and by an unfavorable loss of entropy. A single LNA modification confines the local conformation of nucleotides, causing a smaller, less unfavorable entropic loss when the single strand is restricted to the rigid duplex structure. Additional LNAs adjacent to the initial modification appear to enhance stacking and H-bonding interactions because they increase the enthalpic contributions to duplex stabilization. New nearest-neighbor parameters correctly forecast the positive and negative effects of LNAs on mismatch discrimination. Specificity is enhanced in a majority of sequences and is dependent on mismatch type and adjacent base pairs; the largest discriminatory boost occurs for the central +C.C mismatch within the +T+C+C sequence and the +A.G mismatch within the +T+A+G sequence. LNAs do not affect specificity in some sequences and even impair it for many +G.T and +C.A mismatches. The level of mismatch discrimination decreases the most for the central +G.T mismatch within the +G+G+C sequence and the +C.A mismatch within the +G+C+G sequence. We hypothesize that these discrimination changes are not unique features of LNAs but originate from the shift of the duplex conformation from B-form to A-form.|Base Pair Mismatch[MESH]|Base Sequence[MESH]|DNA/*chemistry[MESH]|Models, Biological[MESH]|Nucleic Acid Conformation[MESH]|Nucleic Acids/*chemistry[MESH]|Oligonucleotides/*chemistry[MESH]|Software[MESH]|Thermodynamics[MESH] |
