![]() ![]() Virus (FMDV), a member of the picornavirus. Small peptide first identified by Ryan and colleagues in the foot-and-mouth disease These limitations can be overcome by a 2A peptide, a “self-cleaving” Lower than that of a gene located before IRES. Second, translation efficiency of a gene placed after the IRES is much When a large insert is cloned into IRES-containing viral vectors with limitedĬloning capacity or when multiple IRESs are used to generate multicistronic The size of IRES is usually longer than 500 nucleotides, which could be a problem Subcellular localization sequences to the gene after IRES and (iii) availability ofĬommercial expression plasmids harboring IRES. Of these strategies, IRES has been widely used due to the following advantages: (i)Įnsured coexpression of genes before and after the IRES (ii) feasibility of adding Proteolytic cleavage sites between genes and (iv) insertion of internal ribosomalĮntry sites (IRESs) between genes. Genes whose expressions are driven by a single promoter (iii) insertion of Reading frames (ORFs) (ii) insertion of splicing signals between genes fusion of Strategies have been employed: (i) multiple promoters fused to the genes' open Organisms using a single plasmid is sometimes required. In biomedical research, the simultaneous expression of more than one gene in cells or All authors confirm adherence toĪll the PLoS ONE policies on sharing data and materials. In study design, data collection and analysis, decision to publish, orĬompeting interests: SRL is an employee of Kim & Jung Co. (R13-2002-0) at Chonnam National University. Health Technology R&D Project, Ministry for Health, Welfare & FamilyĪffairs, Republic of Korea (A100402) and the Korea Science and Engineeringįoundation through the Medical Research Center for Gene Regulation National Research Foundation of Korea (NRF) funded by the Ministry of Education, Terms of the Creative Commons Attribution License, which permits unrestricted use,ĭistribution, and reproduction in any medium, provided the original author andįunding: This work was supported by the Basic Science Research Program through the This is an open-access article distributed under the Received: JanuAccepted: MaPublished: April 29, 2011Ĭopyright: © 2011 Kim et al. PLoS ONE 6(4):Įditor: Volker Thiel, Kantonal Hospital St. (2011) High Cleavage Efficiency of a 2A Peptide Derived from Porcine Teschovirus-1 in Human Cell Lines, Zebrafish and Mice. WeĪnticipate that the 2A-harboring cloning vectors we generated and the highestĮfficiency of the P2A peptide we demonstrated would help biomedical researchersĮasily adopt the 2A technology when bicistronic or multicistronic expression isĬitation: Kim JH, Lee S-R, Li L-H, Park H-J, Park J-H, Lee KY, et al. (P2A) has the highest cleavage efficiency in all the contexts examined. Revealed that among the four 2As, the one derived from porcine teschovirus-1 Western blotting and confocal microscopic analyses Their cleavage efficiency in three commonly used human cell lines, zebrafishĮmbryos and adult mice. ![]() Generated four expression plasmids each harboring different 2A peptides derivedįrom the foot-and-mouth disease virus, equine rhinitis A virus, ThoseaĪsigna virus and porcine teschovirus-1, respectively, and evaluated Reported to date has not been performed in different contexts. No publicly available cloning vectors harboring a 2A peptide gene and (ii)Ĭomprehensive comparison of cleavage efficiency among various 2A peptides Despite theĪdvantages of the 2A peptides, its use is not widespread because (i) there are Good candidate to replace IRES because of its small size and high cleavageĮfficiency between genes upstream and downstream of the 2A peptide. New strategy was required to replace IRES. Due to the large size andĭifference in expression levels between genes before and after IRES, however, a Ribosomal entry site (IRES) has been widely used. Among various strategiesĮmployed to construct bicistronic or multicistronic vectors, an internal Multicistronic expression vectors have been used. ![]() When expression of more than one gene is required in cells, bicistronic or ![]()
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