Part:BBa_K3890002
LP4/2A fusion self cleaving peptide
Fusion of the first 9 aminoacids of the LP4 plant peptide and 20 aminoacids of the F2A viral peptide. This part allows more than one gene to be expressed under the same promoter in eukayotes, by the method of ribosome skipping. When using only the viral peptide, sometimes the process might not be totally succesful, creating non functional proteins. By linking the LP4, the cleaving efficiency can grow up to 80% to 90% [1].
Usage and Biology
Genes co-expression in eukaryotes can be very interesting for many types of studies. It also helps lower the cost and complexity of a genetic circuit. One of the most efficient methods for that is the use of 2A peptides. They are oligopeptides composed of 18–22 amino-acid that are capable of cleaving polypeptides, turning it into two different proteins [2]. They can be found in viruses, being the first one discovered the F2A, from foot- and- mouth disease virus, but there are many more: E2A, T2A, P2A, each with their own cleavage efficiency [2]. And because they are short, they are more guaranteed to not affect the protein function [2]. The mechanism through which they cleave peptides is called ribosome skipping, occuring during translation [2]. But not always the cleaving is totally efficient, producing two separate functional proteins, the ribosome can successfully skip and produce two cleaved proteins, successfully skip but then fall-off and produce only the upstream protein, or not skip, resulting in a fusion protein [2]. That lowers the production of the downstream protein by as much as 50% [2]. But studies have shown an alternative to increase the cleaving efficiency, based on a LP4 peptide. LP4 is a linker peptide from seeds of Impatiens balsamina, capable of spliting between its first and second amino acids [1]. When fusioning the first 9 amino acids of the LP4 to 20 amino acids from the F2A peptide, the efficiency of peptide cleaving has been reported to grow up to 80-90%, guaranteeing a better expression for the downstream protein [1]
References
1.SUN H, Zhou N, Wang H, Huang D, Lang Z. Processing and targeting of proteins derived from polyprotein with 2A and LP4/2A as peptide linkers in a maize expression system. PLoS One. 2017 Mar 30;12(3):e0174804. doi: 10.1371/journal.pone.0174804. PMID: 28358924; PMCID: PMC5373624. 2.LIU, Z., Chen, O., Wall, J.B.J. et al. Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Sci Rep 7, 2193 (2017). https://doi.org/10.1038/s41598-017-02460-2
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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