Optimization of Bicistronic Lentiviral Vector Service
Bicistronic lentiviral vector is a type of newly developed therapeutic gene delivery system with the ability of co-expressing two genes in infected cells. As an undisputed gene therapy service provider, Creative Biolabs offers a full range of bicistronic lentiviral vector construction services based on our powerful GTOnco™ Platform platform and abundant experience.
Introduction of Bicistronic Lentiviral Vector
Generally, a bicistron is termed as two mRNA molecules encoding different peptide chains that are linked together by a short sequence of non-coding proteins. Actually, the DNA fragments encoding these two peptide chains are located in the same transcription unit and share the same starting point and endpoint, which means that the two peptide chains are usually expressed simultaneously. Exactly, the property that co-expression of two or more (multicistron) specific genes can be promisingly applied for many biological fields, especially for gene therapy.
Lentiviral vectors are useful tools for delivering the target genes into the body or modifying the genes in both dividing and non-dividing cells using lentiviruses. The transgene capacity of lentiviral vectors is much larger than common vectors, which makes those lentiviruses the most effective vehicles to transfer large or complex genes, such as bicistron (or multicistron) to cells. Moreover, bicistronic lentiviral vectors technology allows to simultaneously deliver and co-express two proteins in any cell type.
Figure 1. Schematic diagram of HIV-1 based lentiviral vectors. (Chinnasamy, 2006)
Application of Bicistronic Lentiviral Vector
One of the most widely used applications of the bicistronic lentiviral vector is the insertion of a fluorescent protein gene to determine whether the transgene is successfully expressed in transduced cells. Fernández et al. have constructed bicistronic lentiviral vectors using Δ1D/2A and internal ribosome entry site (IRES) elements for the expression of enhanced green fluorescent protein (EGFP) and cerebral dopamine neurotrophic factor (CDNF) to track the expression of CDNF in rats. By these bicistronic lentiviral vectors, they successfully detected the cells stable expressing CDNF by EGFP fluorescence. The fluorescence indicated that CDNF proteins were properly recruited and processed in the endoplasmic reticulum and had an effect on neuronal morphology.
Figure 2. Expression of CDNF by bicistronic vectors in hippocampal neurons. (Fernández, 2014)
Bicistronic lentiviral vector also plays potential roles in cancer gene therapy. Scientists have constructed a bicistronic lentiviral vector using different 2A peptides to express two T-cell receptors that directly against the melanoma differentiation antigens glycoprotein 100 and melanoma-associated antigen recognized by T cells (MART-1). They modified the bicistronic lentiviral vector by inserting amino-acid spacer sequences and a furin cleavage site, by which the novel vector induced an increased expression of T-cell receptors in transduced lymphocytes. This co-expressed bicistronic lentiviral vector exhibited a strong anti-melanoma activity and was potentially used in cancer gene therapy.
Advantages
In terms of co-expressing your genes of interest, Creative Biolabs is a first-rate biotech company who has comprehensive development capability to provide you a wide range of customized gene therapy services. We have absolute advantages in offering lentiviral vector services:
- Customized design of bicistron (or multicistron) based on clients' demands
- High-quality lentiviral vectors construction
- Various fluorescence labels are available
- High-efficiency expression of your genes of interest
For more detailed information, please feel free to contact us or directly send us an online inquiry.
References
- Chinnasamy, D.; et al. (2006). Multicistronic lentiviral vectors containing the FMDV 2A cleavage factor demonstrate robust expression of encoded genes at limiting MOI. Virology Journal. 3:14.
- Fernández, A.; et al. (2014). Construction of bicistronic lentiviral vectors for tracking the expression of CDNF in transduced cells. Plasmid. 76: 15-23.
- Yang, S.; et al. (2008). Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition. Gene Therapy. 15(21): 1411-1423.
