Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (1): 104-108    DOI:
    
Development of 2A Peptide-based Strategies for Constructing Multicistronic Expression Vectors
ZHANG Huan1, HUANG Si-chao2, CAI Shao-hui1
1. College of Pharmacy, Jinan University, Guangzhou 510632, China;
2. Department of Pharmacy, The People’s Hospital of Zhuhai, Zhuhai 519000, China
Download: HTML   PDF(392KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  The construction of multicistronic vectors is important for biotechnology, especially for disease therapy. However, there are some limitations of the commonly used vectors such as limited capcity of vectors, imbalance of protein expression, low activity of protein or limited in the same subcellular space. 2A peptide has been used for construction of multicistronic vectors in recent years. Vectors containing 2A peptide definitely outweigh other construction strategies. The resource of 2A peptide, cleave mechanism, biological character, the relationship with protein targeting and application are reviewed.

Key wordsMulticistronic vectors      2A peptide      Co-expression     
Received: 04 September 2012      Published: 25 January 2013
ZTFLH:  Q782  
Cite this article:

ZHANG Huan, HUANG Si-chao, CAI Shao-hui. Development of 2A Peptide-based Strategies for Constructing Multicistronic Expression Vectors. China Biotechnology, 2013, 33(1): 104-108.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2013/V33/I1/104

[1] Szymczak-Workman A L, Vignali K M,Vignali D A. Design and construction of 2A peptide-linked multicistronic vectors. Cold Spring Harb Protoc, 2012, 2012(2): 199-204.
[2] Li G, Xiang L, Yang W, et al. Efficient multicistronic co-expression of hNIS and hTPO in prostate cancer cells for nonthyroidal tumor radioiodine therapy. Am J Nucl Med Mol Imaging, 2012, 2(4): 483-498.
[3] Mir F A, Kaufmann S H,Eddine A N. A multicistronic DNA vaccine induces significant protection against tuberculosis in mice and offers flexibility in the expressed antigen repertoire. Clin Vaccine Immunol, 2009, 16(10): 1467-1475.
[4] Ralley L, Enfissi E M, Misawa N, et al. Metabolic engineering of ketocarotenoid formation in higher plants. Plant J, 2004, 39(4): 477-486.
[5] Yeo E T, Kwon H B, Han S E, et al. Genetic engineering of drought resistant potato plants by introduction of the trehalose-6-phosphate synthase (TPS1) gene from Saccharomyces cerevisiae. Mol Cells, 2000, 10(3): 263-268.
[6] Deng W, Yang D, Zhao B, et al. Use of the 2A peptide for generation of multi-transgenic pigs through a single round of nuclear transfer. PLoS One, 2011, 6(5): e19986.
[7] Chinnasamy D, Milsom M D, Shaffer J, et al. Multicistronic lentiviral vectors containing the FMDV 2A cleavage factor demonstrate robust expression of encoded genes at limiting MOI. Virol J, 2006, 3: 14.
[8] Szymczak A L, Vignali D A. Development of 2A peptide-based strategies in the design of multicistronic vectors. Expert Opin Biol Ther, 2005, 5(5): 627-638.
[9] de Felipe P. Skipping the co-expression problem: the new 2A "CHYSEL" technology. Genet Vaccines Ther, 2004, 2(1): 13.
[10] Szymczak-Workman A L, Vignali K M,Vignali D A. Generation of 2A-linked multicistronic cassettes by recombinant PCR. Cold Spring Harb Protoc, 2012, 2012(2): 251-254.
[11] Ryan M D, King A M,Thomas G P. Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence. J Gen Virol, 1991, 72 ( Pt 11): 2727-2732.
[12] Donnelly M L, Hughes L E, Luke G, et al. The 'cleavage' activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring '2A-like' sequences. J Gen Virol, 2001, 82(Pt 5): 1027-1041.
[13] Donnelly M L, Luke G, Mehrotra A, et al. Analysis of the aphthovirus 2A/2B polyprotein 'cleavage' mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal 'skip’. J Gen Virol, 2001, 82(Pt 5): 1013-1025.
[14] Sharma P, Yan F, Doronina V A, et al. 2A peptides provide distinct solutions to driving stop-carry on translational recoding. Nucleic Acids Res, 2012, 40(7): 3143-3151.
[15] Doronina V A, Wu C, de Felipe P, et al. Site-specific release of nascent chains from ribosomes at a sense codon. Mol Cell Biol, 2008, 28(13): 4227-4239.
[16] de Felipe P, Luke G A, Hughes L E, et al. E unum pluribus: multiple proteins from a self-processing polyprotein. Trends Biotechnol, 2006, 24(2): 68-75.
[17] Kim J H, Lee S R, Li L H, et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS One, 2011, 6(4): e18556.
[18] Yang S, Cohen C J, Peng P D, et al. Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition. Gene Ther, 2008, 15(21): 1411-1423.
[19] Provost E, Rhee J,Leach S D. Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos. Genesis, 2007, 45(10): 625-629.
[20] Trichas G, Begbie J,Srinivas S. Use of the viral 2A peptide for bicistronic expression in transgenic mice. BMC Biol, 2008, 6: 40.
[21] de Felipe P, Ryan M D. Targeting of proteins derived from self-processing polyproteins containing multiple signal sequences. Traffic, 2004, 5(8): 616-626.
[22] de Felipe P, Luke G A, Brown J D, et al. Inhibition of 2A-mediated 'cleavage’ of certain artificial polyproteins bearing N-terminal signal sequences. Biotechnol J, 2010, 5(2): 213-223.
[23] Yan J, Wang H, Xu Q, et al. Signal sequence is still required in genes downstream of "autocleaving" 2A peptide for secretary or membrane-anchored expression. Anal Biochem, 2010, 399(1): 144-146.
[24] Okita K, Nakagawa M, Hyenjong H, et al. Generation of mouse induced pluripotent stem cells without viral vectors. Science, 2008, 322(5903): 949-953.
[25] Sommer C A, Stadtfeld M, Murphy G J, et al. Induced pluripotent stem cell generation using a single lentiviral stem cell cassette. Stem Cells, 2009, 27(3): 543-549.
[26] Carey B W, Markoulaki S, Hanna J, et al. Reprogramming of murine and human somatic cells using a single polycistronic vector. Proc Natl Acad Sci U S A, 2009, 106(1): 157-162.
[27] Quintarelli C, Vera J F, Savoldo B, et al. Co-expression of cytokine and suicide genes to enhance the activity and safety of tumor-specific cytotoxic T lymphocytes. Blood, 2007, 110(8): 2793-2802.
[28] Wargo J A, Robbins P F, Li Y, et al. Recognition of NY-ESO-1+ tumor cells by engineered lymphocytes is enhanced by improved vector design and epigenetic modulation of tumor antigen expression. Cancer Immunol Immunother, 2009, 58(3): 383-394.
[29] Lengler J, Holzmuller H, Salmons B, et al. FMDV-2A sequence and protein arrangement contribute to functionality of CYP2B1-reporter fusion protein. Anal Biochem, 2005, 343(1): 116-124.
[30] Ibrahimi A, Vande V G, Reumers V, et al. Highly efficient multicistronic lentiviral vectors with peptide 2A sequences. Hum Gene Ther, 2009, 20(8): 845-860.
[31] Ohashi Y, Tsubota T, Sato A, et al. A bicistronic lentiviral vector-based method for differential transsynaptic tracing of neural circuits. Mol Cell Neurosci, 2011, 46(1): 136-147.
[32] Szymczak A L, Workman C J, Wang Y, et al. Correction of multi-gene deficiency in vivo using a single ‘self-cleaving’ 2A peptide-based retroviral vector. Nat Biotechnol, 2004, 22(5): 589-594.
[1] Yuan TIAN,Yan-ling LI. Biosynthesis of Fusaruside Based on Recombinant Pichia pastoris[J]. China Biotechnology, 2019, 39(7): 8-14.
[2] Yin YAO,Qi MIN,Hai-rong XIONG,Li ZHANG. Co-expression of xylanase and mannanase in Pichia pastoris and the enzymatic analyses[J]. China Biotechnology, 2019, 39(3): 37-45.
[3] REN Li-qiong,WU Jing,CHEN Sheng. Co-Expression of N-Acetyltransferase Enhances the Expression of Aspergillus nidulans α-Glucosidase in Pichia pastoris[J]. China Biotechnology, 2019, 39(10): 75-81.
[4] LIANG Dong, XING Yong-qiang, CAI Lu. The Construction and Analysis of the Coexpression Network for the Gene Related to Renal Tumor[J]. China Biotechnology, 2016, 36(2): 30-37.
[5] HE Zhang-hua, WANG Yang, ZHAO Jun, LIU Xiao-jie, ZHANG Li-hua, WANG Dong, SHI Ming-lei, HUANG Fen, YOU Ping, ZHAO Zhi-hu. Construction of a Vector Suitable for the Tandem Coexpression of Multiple Genes by a Single Plasmid[J]. China Biotechnology, 2011, 31(01): 40-45.
[6] QI Xianghui Tian LIANG. Molecular Cloning, Co-expression and Characterization of dhaF and dhaG Genes Encoding Glycerol Dehydratase Reactivating Factor of Citrobacter freundii[J]. China Biotechnology, 2009, 29(01): 39-43.
[7] . Construction of dhaBCE and yqhD co-expression vector and its biotransformation of glycerol[J]. China Biotechnology, 2008, 28(5): 46-51.