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Original article link: HeYuan Bio – Lentiviral Vectors
Lentiviral vectors are a class of viral vectors derived from the human immunodeficiency virus (HIV) and belong to the retrovirus family. Their genome is RNA, with the virulence‑related genes deleted and replaced by heterologous target genes. As pseudotyped viruses, they can integrate exogenous genes into the host genome, enabling stable expression, and they are capable of infecting both dividing and non‑dividing cells.
After the lentiviral genome enters the cell, it is reverse-transcribed into DNA in the cytoplasm, forming the pre-integration complex. Upon entering the nucleus, the DNA integrates into the host genome. The integrated DNA is then transcribed into mRNA, which returns to the cytoplasm to drive expression of the target protein or the production of small RNAs. Lentivirus‑mediated gene expression or small RNA–based silencing is sustained and stable, and it is inherited through cell division along with the host genome (Figure 1).
Figure 1. Process of lentiviral packaging and cell infection.
Lentiviral vectors are characterized by a broad host range, low immunogenicity, large genetic capacity, and the ability to sustain long-term gene expression. They can efficiently transduce a wide variety of cell types, including cultured tumor cells, hepatocytes, cardiomyocytes, neurons, endothelial cells, and stem cells.
Lentiviral infection exhibits integrative properties, enabling the efficient integration of exogenous genes into the host genome and achieving long-term expression. Consequently, stable cell lines can be established for studying gene‑dependent cellular functions. Moreover, T cells engineered with lentiviral vectors are well suited for CAR‑T cell therapy.
Lentivirus Packaging and Purification
Lentiviral packaging is performed using either a three-plasmid or a four-plasmid system. The cell culture supernatant is collected, and the virus is concentrated, purified, and harvested by ultracentrifugation.
Lentivirus Titer Assay
Detection method: Quantitative PCR is used to measure the copy number of exogenous DNA in the cellular genome following infection.
Experimental principle: Lentiviral vectors mediate the integration of exogenous genes into the target cell genome via reverse transcription.
Advantages of Lentiviral Vectors
① Prolonged expression: Lentiviruses can integrate exogenous genes into the host cell genome, enabling long-term, stable gene expression that is maintained through successive cell divisions and generations. This makes them the preferred choice for cell-based experiments and a common tool for generating stable cell lines.
② High safety: No pathogenicity has been observed; lentiviral vector–engineered T cells are used in CAR‑T cell therapy.
③ Low immunogenicity: Direct injection into living tissues is unlikely to elicit an immune response, making it suitable for animal experiments.
Comparison of the Biological Characteristics of Different Viruses
Carrier Selection
HeYuan Bio offers a comprehensive portfolio of lentiviral products for manipulating both coding and non-coding genes, including lncRNAs, miRNAs, and circRNAs. The following lentiviral vectors are available for your selection (this list is not exhaustive):
|
Regulation method |
Carrier Number |
Vector name (vector element order) |
Eukaryotic resistance |
Fluorescence |
Promoter |
Default label |
Carrier capacity |
|
Overexpression |
GL127 |
pSLenti-CMV-EGFP-3xFLAG-WPRE |
N/A |
EGFP |
CMV |
3FLAG |
4.8kb |
|
GL121 |
pSLenti-EF1-EGFP-CMV-MCS-WPRE |
N/A |
EGFP |
CMV |
N/A |
4.4kb |
|
|
GL129 |
pSLenti-CMV-mCherry-3xFLAG-WPRE |
N/A |
mCherry |
CMV |
3FLAG |
4.8kb |
|
|
GL119 |
pSLenti-CMV-MCS-3xFLAG-PGK-Puro-WPRE |
Pure |
N/A |
CMV |
3FLAG |
4.4kb |
|
|
GL120 |
pSLenti-SFH-EGFP-P2A-Puro-CMV-MCS-3xFLAG-WPRE |
Pure |
EGFP |
CMV |
3FLAG |
3.5kb |
|
|
GL107 |
pSLenti-EF1-EGFP-P2A-Puro-CMV-MCS-3xFLAG-WPRE |
Pure |
EGFP |
CMV |
3FLAG |
3.7kb |
|
|
GL109 |
pSLenti-EF1-EGFP-F2A-Puro-CMV-MCS-WPRE |
Pure |
EGFP |
CMV |
N/A |
3.8kb |
|
|
GL122 |
pSLenti-EF1-EGFP-F2A-BSR-CMV-MCS-WPRE |
blasticidin |
EGFP |
CMV |
N/A |
3.9kb |
|
|
GL130 |
pSLenti-CMV-EGFP-3xFLAG-PGK-Puro-WPRE |
Pure |
EGFP |
CMV |
3FLAG |
3.7kb |
|
|
GL132 |
pSLenti-EF1-EGFP-F2A-Puro-WPRE2-CMV-MCS |
Pure |
EGFP |
CMV |
N/A |
3.7kb |
|
|
GL123 |
pSLenti-EF1-mCherry-P2A-Puro-CMV-MCS-3xFLAG-WPRE |
Pure |
mCherry |
CMV |
3FLAG |
3.7kb |
|
|
GL125 |
pSLenti-CMV-mCherry-3xFLAG-PGK-Puro-WPRE |
Pure |
mCherry |
CMV |
3FLAG |
3.7kb |
|
|
GL133 |
pSLenti-EF1-mCherry-F2A-Puro-WPRE2-CMV-MCS |
Pure |
mCherry |
CMV |
N/A |
3.7kb |
|
|
H114 |
pLenti-CMV-Luc2-IRES-Puro-WPRE |
Pure |
N/A |
CMV |
N/A |
2.2kb |
|
|
GL124 |
pSLenti-EF1-Luc2-F2A-Puro-CMV-MCS-WPRE |
Pure |
N/A |
CMV |
N/A |
2.8kb |
|
|
Express cre |
H126 |
pLenti-CMV-NLS-Cre-3xFLAG-WPRE |
N/A |
N/A |
CMV |
3FLAG |
3.9kb |
|
H15108 |
pLenti-CMV-DIO-MCS-WPRE |
N/A |
N/A |
CMV |
3FLAG |
5.3kb |
|
|
Three Standards |
H7656 |
pLenti-CBh-3xFLAG-Luc2-tCMV-mNeonGreen-F2A-Puro-WPRE |
Pure |
mNeonGreen |
CBh |
3FLAG |
1.3kb |
|
H7657 |
pLenti-CBh-3xFLAG-Luc2-tCMV-tdTomato-F2A-Puro-WPRE |
Pure |
tdTomato |
CBh |
3FLAG |
0.6kb |
|
|
H9911 |
pLenti-CBh-3xFLAG-Luc2-tCMV-mNeonGreen-F2A-BSR-WPRE |
blasticidin |
mNeonGreen |
CBh |
3FLAG |
1.5kb |
|
|
H9912 |
pLenti-CBh-3xFLAG-Luc2-tCMV-tdTomato-F2A-BSR-WPRE |
blasticidin |
tdTomato |
|
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