- [Protocol] Development of DNA aptamers specific for small therapeutic peptides using a modified SELEX method
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Jaemin Lee , Minkyung Ryu , Dayeong Bae , Hong-Man Kim , Seong-il Eyun , Jeehyeon Bae , Kangseok Lee
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J. Microbiol. 2022;60(7):659-667. Published online June 22, 2022
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DOI: https://doi.org/10.1007/s12275-022-2235-4
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Abstract
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Aptamers are short single-stranded DNA or RNA oligonucleotides
capable of binding with high affinity and specificity
to target molecules. Because of their durability and ease of synthesis,
aptamers are used in a wide range of biomedical fields,
including the diagnosis of diseases and targeted delivery of
therapeutic agents. The aptamers were selected using a process
called systematic evolution of ligands by exponential enrichment
(SELEX), which has been improved for various research
purposes since its development in 1990. In this protocol,
we describe a modified SELEX method that rapidly produces
high aptamer screening yields using two types of magnetic
beads. Using this method, we isolated an aptamer that
specifically binds to an antimicrobial peptide. We suggest that
by conjugating a small therapeutic-specific aptamer to a gold
nanoparticle-based delivery system, which enhances the stability
and intracellular delivery of peptides, aptamers selected
by our method can be used for the development of therapeutic
agents utilizing small therapeutic peptides.
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Citations
Citations to this article as recorded by 
- Recent approaches in the application of antimicrobial peptides in food preservation
Satparkash Singh, Bhavna Jha, Pratiksha Tiwari, Vinay G. Joshi, Adarsh Mishra, Yashpal Singh Malik World Journal of Microbiology and Biotechnology.2024;[Epub] CrossRef - Design and application of microfluidics in aptamer SELEX and Aptasensors
Shikun Zhang, Yingming Zhang, Zhiyuan Ning, Mengxia Duan, Xianfeng Lin, Nuo Duan, Zhouping Wang, Shijia Wu Biotechnology Advances.2024; 77: 108461. CrossRef - Nanogenosensors based on aptamers and peptides for bioelectrochemical cancer detection: an overview of recent advances in emerging materials and technologies
Babak Mikaeeli Kangarshahi, Seyed Morteza Naghib Discover Applied Sciences.2024;[Epub] CrossRef - Recent progress of SELEX methods for screening nucleic acid aptamers
Chao Zhu, Ziru Feng, Hongwei Qin, Lu Chen, Mengmeng Yan, Linsen Li, Feng Qu Talanta.2024; 266: 124998. CrossRef - Aptamer-conjugated gold nanoparticles platform as the intracellular delivery of antibodies for cancer therapy
Ji-Hyun Yeom, Eunkyoung Shin, Hanyong Jin, Haifeng Liu, Yongyang Luo, Youngwoo Nam, Minkyung Ryu, Wooseok Song, Heeyoun Chi, Jeongkyu Kim, Kangseok Lee, Jeehyeon Bae Journal of Industrial and Engineering Chemistry.2023; 126: 480. CrossRef - Regulation of transforming growth factor-β signaling as a therapeutic approach to treating colorectal cancer
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- Functional Analysis of the Invariant Residue G791 of Escherichia coli 16S rRNA
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Woo-Seok Song , Hong-Man Kim , Jae-Hong Kim , Se-Hoon Sim , Sang-Mi Ryou , Sanggoo Kim , Chang-Jun Cha , Philip R. Cunningham , Jeehyeon Bae , Kangseok Lee
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J. Microbiol. 2007;45(5):418-421.
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DOI: https://doi.org/2595 [pii]
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Abstract
PDF
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The nucleotide at position 791(G791) of E. coli 16S rRNA was previously identified as an invariant residue for ribosomal function. In order to characterize the functional role of G791, base substitutions were introduced at this position, and mutant ribosomes were analyzed with regard to their protein synthesis ability, via the use of a specialized ribosome system. These ribosomal RNA mutations attenuated the ability of ribosomes to conduct protein synthesis by more than 65%. A transition mutation (G to A) exerted a moderate effect on ribosomal function, whereas a transversion mutation (G to C or U) resulted in a loss of protein synthesis ability of more than 90%. The sucrose gradient profiles of ribosomes and primer extension analysis showed that the loss of protein-synthesis ability of mutant ribosomes harboring a base substitution from G to U at position 791 stems partially from its inability to form 70S ribosomes. These findings show the involvement of the nucleotide at position 791 in the association of ribosomal subunits and protein synthesis steps after 70S formation, as well as the possibility of using 16S rRNA mutated at position 791 for the selection of second-site revertants in order to identify ligands that interact with G791 in protein synthesis.
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