LNA (Locked Nucleic Acid)

LNA (Locked Nucleic Acid)

What is an LNA? A locked nucleic acid (LNA), often referred to as inaccessible RNA, is a modified RNA nucleotide. The ribose moiety of an LNA nucleotide is modified with an extra bridge connecting the 2′ oxygen and 4′ carbon. The bridge “locks” the ribose in the 3′-endo (North) conformation, which is often found in the A-form duplexes. LNA nucleotides can be mixed with DNA or RNA residues in the oligonucleotide whenever desired and hybridize with DNA or RNA according to Watson-Crick base-pairing rules. Such oligomers are synthesized chemically and are commercially available. The locked ribose conformation enhances base stacking and backbone pre-organization. This significantly increases the hybridization properties (melting temperature) of oligonucleotides. LNA was independently synthesized by the group of Jesper Wengel in 1998, soon after the first synthesis by the group of Takeshi Imanishi in 1997. The exclusive rights to the LNA technology were secured in 1997 by Exiqon A/S, a Danish biotech company.

LNA nucleotides are used to increase the sensitivity and specificity of expression in DNA microarrays, FISH probes, quantitative PCR probes and other molecular biology techniques based on oligonucleotides. For the in situ detection of miRNA, the use of LNA is currently (2005) the only efficient method. A triplet of LNA nucleotides surrounding a single-base mismatch site maximizes LNA probe specificity unless the probe contains the guanine base of G-T mismatch.

Using LNA-based oligonucleotides therapeutically is an emerging field of biotechnology. The Danish pharmaceutical company Santaris Pharma a/s owns the sole rights to therapeutic uses of LNA technology and is now developing a new, LNA-based, hepatitis C drug called miravirsen, targeting miR-122, which is in Phase II clinical testing as of late 2010.

Definition of an LNA?

Locked nucleic acid (LNA) is a nucleic acid analogue containing one or more LNA nucleotide monomers with a bicyclic furanose unit locked in an RNA mimicking sugar conformation. LNA oligonucleotides display unprecedented hybridization affinity toward complementary single-stranded RNA and complementary single- or double-stranded DNA. Structural studies have shown that LNA oligonucleotides induce A-type (RNA-like) duplex conformations. The wide applicability of LNA oligonucleotides for gene silencing and their use for research and diagnostic purposes are documented in a number of recent reports, some of which are described herein.

What is an LNA?

LNA (Locked Nucleic Acids) are synthetic modified nucleic acids where the carbohydrate part of the nucleic acid has been synthetically changed. The modification results in an increased bonding strength between the DNA-bases in a double-helix when one of the DNA-bases has been modified. The overall result is a higher melting point of a DNA double-helix containing LNA-modified nucleic acids and thereby an increased stability. By designing the complementary DNA-strand in a double helix so it consists more or less of LNA-units, it is possible to regulate the rate of transcription – even to block it completely. In this way, it is possible to control the expression of genes and thereby the synthesis of selected proteins. The LNA technology is, therefore, a promising tool in the treatment of diseases which originate from genetic defects.