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

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.

[1]  Weihong Tan,et al.  Engineering DNA aptamers for novel analytical and biomedical applications , 2011 .

[2]  Subash C. B. Gopinath,et al.  Prospects of Ligand-Induced Aptamers , 2008 .

[3]  Subash C. B. Gopinath,et al.  Waveguide-Mode Sensors as Aptasensors , 2012, Sensors.

[4]  Koichi Awazu,et al.  Colorimetric detection of controlled assembly and disassembly of aptamers on unmodified gold nanoparticles , 2013, Biosensors and Bioelectronics.

[5]  David R. Latulippe,et al.  RAPID-SELEX for RNA Aptamers , 2013, PLoS ONE.

[6]  Subash C B Gopinath,et al.  An RNA aptamer that distinguishes between closely related human influenza viruses and inhibits haemagglutinin-mediated membrane fusion. , 2006, The Journal of general virology.

[7]  A Mukherjee,et al.  Nucleic acid aptamers: clinical applications and promising new horizons. , 2011, Current medicinal chemistry.

[8]  Catherine Lozupone,et al.  Selection of the simplest RNA that binds isoleucine. , 2003, RNA.

[9]  Subash C. B. Gopinath,et al.  Generation of anti-influenza aptamers using the systematic evolution of ligands by exponential enrichment for sensing applications. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[10]  S. C. B. Gopinath Antiviral aptamers , 2007, Archives of Virology.

[11]  E. Peyrin,et al.  Macrocyclic host-dye reporter for sensitive sandwich-type fluorescent aptamer sensor. , 2015, Analytical chemistry.

[12]  S. Gopinath Methods developed for SELEX , 2006, Analytical and bioanalytical chemistry.

[13]  Ivet Bahar,et al.  Functional motions of influenza virus hemagglutinin: a structure-based analytical approach. , 2002, Biophysical journal.

[14]  S. Gopinath,et al.  Anti-coagulant aptamers. , 2008, Thrombosis research.

[15]  Subash C. B. Gopinath,et al.  Aptamer That Binds to the gD Protein of Herpes Simplex Virus 1 and Efficiently Inhibits Viral Entry , 2012, Journal of Virology.

[16]  Makoto Fujimaki,et al.  A high-performance waveguide-mode biosensor for detection of factor IX using PEG-based blocking agents to suppress non-specific binding and improve sensitivity. , 2013, The Analyst.