Shortening full-length aptamer by crawling base deletion – Assisted by Mfold web server application

Subash C.B. Gopinath , Thangavel Lakshmipriya , M.K. Md Arshad , C.H. Voon , Tijjani Adam , Uda Hashim , Harbant Singh , Suresh V. Chinni
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引用次数: 13

Abstract

Systematic Evolution of Ligands by EXponential enrichment (SELEX) is the method to select the specific aptamer against a wide range of targets. For this process, the initial library usually has a length of random sequences from ∼25 and it reaches over 100 bases. The lengthy sequences have disadvantages such as difficult to prepare, less stable and expensive. It is wise to prefer shorter version of aptamer for a wide range of applications including drug delivery process. It is a common practice to shorten the full-length aptamer by mapping analyses and it is tedious. Here, we used a crawling method to shorten the aptamer by different sequential deletion of bases from both 5′ and 3′ ends, assisted by Mfold web server application. Two different kinds of aptamer with varied lengths (randomized region of 30 and 74 bases) were desired for this study, generated against Influenza A/Panama/2007/1999 (H3N2) and gD protein of Herpes Simplex Virus-1. It was found that shortening the aptamer length by crawling pattern is possible with the assistance of Mfold web server application. The obtained results resemble the shortened aptamer derived by mapping analyses. The proposed strategy is recommended to predict the shorter aptamer without involving any wet experimental section.

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通过爬行碱基删除缩短全长适体- Mfold web服务器应用程序辅助
指数富集配体的系统进化(SELEX)是一种针对广泛靶标选择特定适配体的方法。在这个过程中,初始文库的随机序列长度通常在~ 25之间,达到100多个碱基。冗长的序列具有制备困难、稳定性差和价格昂贵等缺点。对于包括给药过程在内的广泛应用而言,选择较短版本的适体是明智的。通过映射分析来缩短全长适配体是一种常见的做法,这是乏味的。在Mfold web server应用程序的辅助下,我们采用爬行法从5 '端和3 '端不同顺序地删除碱基来缩短适配体。本研究需要两种不同长度的适体(30和74个碱基的随机区域),分别针对甲型流感/巴拿马/2007/1999 (H3N2)和单纯疱疹病毒-1的gD蛋白产生。发现在Mfold web服务器应用程序的帮助下,通过爬行模式缩短适配体长度是可能的。所得结果与通过图谱分析得到的缩短的适体相似。所提出的策略被推荐用于预测较短的适体,而不涉及任何湿实验切片。
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