Mengenal Tentang PNA dan Perbedaannya Dengan DNA dari Panagene

Apa itu PNA?

Pertama kali ditemukan oleh Profesor Nielsen, Egholm, Berg dan Buchardt dari University of Copenhagen, Denmark pada tahun 1991.

PNA merupakan kepanjangan dari Peptide Nucleic Acid, merupakan sebuah analog DNA yang dibuat secara artifisial.

PNA memiliki struktur di mana gugus fosfat-ribosa dari DNA disubstitusi dengan gugus amida seperti peptida (N- (2-aminoetil) glisin). Jadi afinitas pengikatan dan stabilitas pada DNA target atau RNA sangat meningkat. Meskipun terjadi perubahan struktural pada gugus tersebut, PNA dapat digunakan dalam berbagai aplikasi di mana DNA dapat digunakan karena dapat membuat ikatan komplementer dengan urutan target seperti halnya DNA.

FEATURES & ADVANTAGES

  • High binding affinity

PNA has electrically neutral amide backbone. There is no repulsion between the PNA strand and the DNA strand, and PNA has the stronger binding affinity for the target DNA.

  • High specificity and sensitivity

The difference in binding affinity between perfect match andmismatch of PNA/DNA strands is bigger than DNA/DNA strands. It is easy to discriminate even single nucleotide mismatched sequence.

  • Chemical/Biological/thermal stability

PNA is quite stable under high temperature or high pH condition. Also, PNA backbone is completely different from DNA’s or RNA’s andis similar to but slightly different from the peptide backbone.So, PNA is stable against enzymes as nuclease or protease.

 

  • Independence against salt in hybridization

PNA has stable and strong binding affinity due to itsindependence against high salt concentration in hybridization.

PNA VS DNA

Feature PNA DNA
Binding Affinity At Least 1°C Higher Tm per Base
Hybridization rate 100-5000 Times Faster
Salt concentration in hybridization Independent Dependent
ΔTm for single nucleotide mismatch <10°C <15°C
ChemicalStability Stable Unstable in Acidic or Basic condition
BiologicalStability Enzyme-Resistant Degradable by Nuclease
Solubility Moderate(but can be improved by simple modification) Good
General Probe Length 13-18mer 25-30mer

 

PANAGENE’s PNA

PANAGENE has developed its proprietary Bts PNA monomers (Bts; benzothiazole-2-sulfonyl group) and PNA oligo synthesis method using Bts monomers, and has supplied high quality PNA oligos.

Bts group of the Bts PNA monomers not only protects the amine group (NH2) of the monomer but also is self-activated. So, oligo synthesis using Bts monomers does not require a pre-activation step or many coupling reagents. Unlike the conventional synthesis method (using Fmoc PNA monomers), the side reactions such as transacylation which occur during the deprotection process are minimized and mass production of high purity PNA oligo can be achieved at low cost. In addition, it is more economical because it is possible to recover and reuse excess Bts monomers which remains after the coupling reaction.

  • Chemical Structure of Bts PNA monomer & oligomerization cycle

[Reference]

Hyunil Lee et al. 2007. Peptide Nucleic Acid Synthesis by Novel Amide Formation. Org. Lett. 9 (17), 3291-3293.

 

  • Bts vs. Fmoc
Type Bts monomer Fmoc monomer
Coupling reagent Not necessary HATU
Mass production Easy Difficult
Side reaction <1% Hard to control(5~10%)
Purification Easy Difficult
Yield >80% <40%
Cost of synthesis Low High

 

Fmoc monomer 


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