Turner John J, Williams Donna, Owen David, Gait Michael J
Medical Research Council, Laboratory of Molecular Biology, Cambridge, United Kingdom.
Curr Protoc Nucleic Acid Chem. 2006 Apr;Chapter 4:Unit 4.28. doi: 10.1002/0471142700.nc0428s24.
Peptide conjugation of oligonucleotides and their analogs is being studied widely towards improving the delivery of oligonucleotides into cells. Amongst the many possible routes of conjugation, the disulfide linkage has proved to be the most popular. This reversible linkage may have advantages for cell delivery, since it is likely to be cleaved within cells, thus releasing the oligonucleotide cargo. It is straightforward to introduce thiol functionalities into both oligonucleotide and peptide components suitable for disulfide conjugation. However, severe difficulties have been encountered in carrying out conjugations between highly cationic peptides and negatively charged oligonucleotides because of aggregation and precipitation. Presented here are reliable protocols for disulfide conjugation that have been verified for both cationic and hydrophobic peptides as well as oligonucleotides containing deoxyribonucleosides, ribonucleosides, 2'-O-methylribonucleosides, locked nucleic acid (LNA) units, as well as phosphorothioate backbones. Also presented are reliable protocols for disulfide conjugation of peptide nucleic acids (PNAs) with peptides.
为了提高寡核苷酸向细胞内的递送效率，寡核苷酸及其类似物的肽缀合正在被广泛研究。在众多可能的缀合途径中，二硫键已被证明是最常用的。这种可逆键可能对细胞递送具有优势，因为它可能在细胞内被裂解，从而释放寡核苷酸负载。将硫醇官能团引入适合二硫键缀合的寡核苷酸和肽组分中很简单。然而，由于聚集和沉淀，在进行高阳离子肽与带负电荷的寡核苷酸之间的缀合时遇到了严重困难。本文介绍了已针对阳离子肽和疏水肽以及含有脱氧核糖核苷、核糖核苷、2'-O-甲基核糖核苷、锁核酸（LNA）单元以及硫代磷酸酯主链的寡核苷酸验证的二硫键缀合可靠方案。还介绍了肽核酸（PNA）与肽的二硫键缀合可靠方案。
