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Lentiviral vectors are powerful tools to a wide range of applications:
In this group, you will find the vectors carrying reporter transgenes, such as GFP or Luciferase. These transgenes are very usefull as control vectors, especially GFP vectors. They can also be used to perfect a transduction protocol by facilitating the expression measurement. You can also find selection transgenes such as neomycin. Such transgenes are very handy when generating a clonal cell line. Vectors in this category are provided with fundamental integrase and envelope (IN(WT) and VSV) to allow a wide range of use.
GFP — Enhanced green fluorescent protein
GFP cDNA encodes a green fluorescent protein (sequence from Aequorea victoria genome, AAB02574.1, 717 pb). This fluorescent reporter protein allows monitoring expression and transduction by fluorescent microscopy or by flow cytometry.
hrGFPII — Humanized recombinant green fluorescent protein
hrGFPII-1 cDNA encodes a humanized recombinant green fluorescent protein (sequence from Renilla reniformis genome, N.R., 717 pb). This fluorescent reporter protein allows monitoring expression and transduction by fluorescent microscopy or by flow cytometry.
LUC — Firelfy luciferase PpyLuc 1
Luc cDNA encodes the firefly luciferase PpyLuc1 (sequence from Photinus pyralis genome, AB644228.1, 1653 pb). This reporter enzyme allows monitoring expression and transduction by bioluminescence assay.
Neo cDNA encodes the neomycin phosphotransferase (sequence from Escherichia coli genome, DD224401.1, 795 pb). This enzyme is responsible for inactivation of aminoglycoside antibiotics (e.g. kanamycin, neomycin, geneticin). It is used as a selection marker and allows, for instance, to create genetically engineered cell lines for in vitro assays.
RFP — Red fluorescent protein
RFP cDNA which encodes red fluorescent protein (artificial sequence derived from sea anemone Entacmaea quadricolor, KT358727.1, 693 pb). RFP is a fluorescent reporter protein allowing the monitoring of expression and transduction by fluorescent microscopy or by flow cytometry.
Vectors sorted in this category are vectors pseudotyped with the Mokola envelope, conferring them a glial target specificity. You can also find vectors whose transgene is placed under control of a cell specific promoter.
MOK — Envelope Glycoprotein from Mokola virus
Mokola is the envelope glycoprotein from the Lyssavirus Mokola. Lentiviral vectors produced with this envelope are expected to be glial cell specific in vivo.
ARR3lg — Arrestin 3 long
Arr3long is the arrestin 3 promoter, long (sequence from pig genome, CU463005.6, 724 pb). This promoter region is expected to drive expression specifically in rod photoreceptor.
ARR3sh — Arrestin 3 short
Arr3short is the arrestin 3 promoter, short (sequence from pig genome, CU463005.6, 249 pb). This promoter region is expected to drive expression specifically in rod photoreceptor.
CaMKII — Fms-like tyrosine kinase-1
CaMKII is the Ca2+/calmodulin-dependent protein kinase II promoter. This promoter region is expected to drive expression specifically in neuronal cells.
Flt1 — Fms-like tyrosine kinase-1
Flt1 is the fms-like tyrosine kinase-1 promoter (sequence from human genome, NG_012003.1, 1036 pb). This promoter region is expected to drive expression specifically in endothelial cells.
GFAP — Glial fibrillary acidic protein
GFAP is the glial fibrillary acidic protein promoter (sequence from human genome, M67446.1, 1674 pb). This promoter region is expected to drive expression specifically in glial cells.
NSE — Neuron specific enolase
NSE is the neuron specific enolase promoter (sequence from rat genome, AB038993.1, 1682 pb). This promoter region is expected to drive expression specifically in neuronal cells.
RHO — Rhodopsin
Rho is the rhodopsin promoter (sequence from mouse genome, AC142099.3, 3868 pb). This promoter region is expected to drive expression specifically in rod photoreceptor.
SYN — Synapsin
SYN is the synapsin promoter (sequence from human genome, NG_008437.1, 469 pb). This promoter region is expected to drive expression specifically in neuronal cells.
Among the great variety of proteins that can be expressed via Lenti-ONE™, you can find proteins with DNA editing function, including Cas9. They can be used to modify the target cell DNA by entering a chosen sequence.
Cre-NLS — Cyclization recombinase fused with nuclear localization signal
Cre-NLS cDNA encodes a cyclization recombinase fused with a nuclear localization signal (sequence from Enterobacterio phage P1 genome, AB449974.1/AX343218.1, 1053 pb). This tyrosine recombinase recognizes LoxP sequences and mediates recombination between two DNA fragments. This reaction can be used to excise a DNA fragment flanked by 2 loxP sequences or to integrate a DNA fragment into a single loxP sequence. Addition of a nuclear localization signal increases the efficiency of the enzyme.
PhiC31-NLS — Integrase phiC31 fused with a nuclear localization signal
PhiC31-NLS cDNA encodes a PhiC31 phage recombinase fused with a nuclear localization signal (sequence from Streptomyces phage PhiC31 genome, AJ006589.2, 1836 pb). This serine recombinase recognizes an attB (or pseudo attB) sequence and an attP (or pseudo-attP) sequence and mediates recombination between both sequences. This allows, for instance, integrating a DNA fragment of interest into host cell chromosomes.
Cas9 — Cas9 cDNA encodes for protein-9 nuclease from Streptococcus pyogenes. (Sequence, WP_010922251.1, 4128bp). Coexpression of Cas9 with an artificial single guide RNA (sgRNA) which fuses the crRNA with the tracrRNA offers the possibility to target and modify DNA sequences of interest.
Cas9-2A-GFP — Cas9 cDNA encodes for protein-9 nuclease from Streptococcus pyogenes. (Sequence, WP_010922251.1, 4128bp). Coexpression of Cas9 with an artificial single guide RNA (sgRNA) which fuses the crRNA with the tracrRNA offers the possibility to target and modify DNA sequences of interest.
GFP cDNA encodes a green fluorescent protein (sequence from Aequorea victoria genome, AAB02574.1, 717 pb). This fluorescent reporter protein allows monitoring expression and transduction by fluorescent microscopy or by flow cytometry.
This group of vectors contains a large amount of vectors. First you can find Lenti-ONE™ with various transgenes driven by promoters specific to the Neurology and/or Ophthalmology areas, such as NSE or Synapsin. You can also fin vectors carrying transgenes related to the Neurology or Ophthalmology areas, such as GDNF or GUCY2Dmut.
BDNF — Brain-derived neurotrophic factor
BDNF cDNA encodes the brain-derived neurotrophic factor isoform a (sequence from human genome, NM_001143805.1/CCDS7866.1, 744 pb). This neurotrophic factor may be used to promote neuronal survival in experimental gene therapy assays.
CNTF — Ciliary neurotrophic factor
CNTF cDNA encodes the ciliary neurotrophic factor (sequence from human genome, NM_000614.3/CCDS 31554.1, 603 pb). This neurotrophic factor may be used to promote neuronal survival in experimental gene therapy assays.
GDNF — Glial cell derived neurotrophic factor
GDNF cDNA encodes the glial cell derived neurotrophic factor (sequence from human genome, BC128108.1, 636 pb). This neurotrophic factor may be used to promote neuronal survival in experimental gene therapy assays.
GPx — Glutathione peroxidase 1
GPX1 cDNA encodes the glutathione peroxidase 1 (sequence from human genome, NM_000581.2/CCDS 43091.1, 612 pb). This peroxidase enzyme is responsible for reducing hydrogen peroxide to water.
GUCY2Dmut — Guanylate cyclase 2D
GUCY2Dmut cDNA encodes a mutant guanylate cyclase 2D with two substitutions: 837 D>E, 838 R>S (modified sequence from human genome, NM_000180.3, 3641 pb). This enzyme is guanylate cyclase 2D with dominant negative substitution D837E and R838S. These mutations have been associated in humans with cone dystrophies.
SOD — Superoxide dismutase 1
SOD cDNA encodes the superoxide dismutase 1 (sequence from human genome, NM_000454.4/CCDS 33536.1, 465 pb). This metalloproteinase is responsible for the dismutation of superoxides.
VEGF121 — Vascular endothelial growth factor 121
VEGF121 cDNA encodes the vascular endothelial growth factor 121 (sequence from human genome, AF214570.1, 444 pb). This protein is a growth factor. It favors angiogenesis. It may also promote neuronal survival in experimental gene transfer assays.
VEGF165 — Vascular endothelial growth factor 165
VEGF165 cDNA encodes the vascular endothelial growth factor 165 (sequence from human genome, AF486837.1, 576 pb). This protein is a growth factor. It favors angiogenesis. It may also promote neuronal survival in experimental gene transfer assays.
VEGF165b — Vascular endothelial growth factor 165b
VEGF165b cDNA encodes the vascular endothelial growth factor 165b (sequence from human genome, AF430806.1, 606 pb). This protein is an antagonist of VEGF165 and inhibits angiogenesis process. It may be used in anti-angiogenesis experimental gene transfer assays.
MOK — Envelope Glycoprotein from Mokola virus
Mokola is the envelope glycoprotein from the Lyssavirus Mokola. Lentiviral vectors produced with this envelope are expected to be glial cell specific in vivo.
Optogenetics
Optogenetics is the combination of genetic and optical methods to achieve gain or loss of function of well-defined events in specific cells of living tissue. This method allows staining, enhancing or decreasing the activity of neural, retinal or cardiac cell populations just in using light.
ChR2-2A-GFP — ChR2 cDNA encodes for Channelrhodopsin-2 from Chlamydomonas reinhardtii (NCBI Reference Sequence: XP_001701725.1, 927bp). ChR2 is used in optogenetics for light activation of neural cells. GFP cDNA encodes a green fluorescent protein (sequence from Aequorea victoria genome, AAB02574.1, 717bp). This fluorescent reporter protein allows the monitoring of expression and transduction by fluorescent microscopy or by flow cytometry.
eNpHR3-2A-GFP — eNpHR3.0 cDNA is the engineered form of NpHR, Halorhodopsin from Natronomonas (GenBank: EF474018.1, 876bp). eNpHR3.0 is used in optogenetics for light inhibition of neural cells. GFP cDNA encodes a green fluorescent protein (sequence from Aequorea victoria genome, AAB02574.1, 717 bp). This fluorescent reporter protein allows the monitoring of expression and transduction by fluorescent microscopy or by flow cytometry.
The vectors in this group carry transgenes expressing proteins that can be useful in the study of apoptosis, such as Bcl2, or in the study of oxidative stress, such as AOX. These vectors are available with multiple promoters, integrases and envelopes.
AOX — Alternative oxidase
AOX cDNA encodes an alternative oxidase (sequence from Ciona intestinalis genome, DM193474.1, 1110 pb). This alternative oxidase is part of the electron transport chain in mitochondria. It contributes to reduction of ROS production when overexpressed in mammalian cells.
Bcl2 — B-cell CLL/lymphoma 2
Bcl2 cDNA encodes the B-cell CLL lymphoma 2 isoform alpha (sequence from human genome, NM_000633.2/CCDS 11981.1, 720 pb). This protein is an apoptosis regulating factor.
GPx — Glutathione peroxidase 1
GPX1 cDNA encodes the glutathione peroxidase 1 (sequence from human genome, NM_000581.2/CCDS 43091.1, 612 pb). This peroxidase enzyme is responsible for reducing hydrogen peroxide to water.
SOD — Superoxide dismutase 1
SOD cDNA encodes the superoxide dismutase 1 (sequence from human genome, NM_000454.4/CCDS 33536.1, 465 pb). This metalloproteinase is responsible for the dismutation of superoxides.
In this category you can find vectors whose transgene is under control of a cell specific promoter useful in the field of angiogenesis. But there are also vectors expressing different angiogenic factors. These vectors are available with different integrases, so you can choose the expression kinetic according to your project.
Flt1 — Fms-like tyrosine kinase-1
Flt1 is the fms-like tyrosine kinase-1 promoter (sequence from human genome, NG_012003.1, 1036 pb). This promoter region is expected to drive expression specifically in endothelial cells.
VEGF121 — Vascular endothelial growth factor 121
VEGF121 cDNA encodes the vascular endothelial growth factor 121 (sequence from human genome, AF214570.1, 444 pb). This protein is a growth factor. It favors angiogenesis. It may also promote neuronal survival in experimental gene transfer assays.
VEGF165 — Vascular endothelial growth factor 165
VEGF165 cDNA encodes the vascular endothelial growth factor 165 (sequence from human genome, AF486837.1, 576 pb). This protein is a growth factor. It favors angiogenesis. It may also promote neuronal survival in experimental gene transfer assays.
VEGF165b — Vascular endothelial growth factor 165b
VEGF165b cDNA encodes the vascular endothelial growth factor 165b (sequence from human genome, AF430806.1, 606 pb). This protein is an antagonist of VEGF165 and inhibits angiogenesis process. It may be used in anti-angiogenesis experimental gene transfer assays.
In this section are regrouped vectors with transgenes expressing differentiation factors, like MyoD. Besides, there are also vectors whose transgene in under the control of a cell type-specific promoter. These last can be useful to characterize differentiated cells.
MyoD — Myogenic differentiation
MyoD cDNA encodes a myogenic differentiation factor (sequence from mouse genome, NM_010866.2/CCDS 21277.1, 957 pb). This protein is a myogenic factor which can be used to promote differentiation of stem cells into myotubes.
Flt1 — Fms-like tyrosine kinase-1
Flt1 is the fms-like tyrosine kinase-1 promoter (sequence from human genome, NG_012003.1, 1036 pb). This promoter region is expected to drive expression specifically in endothelial cells.
SYN — Synapsin
SYN is the synapsin promoter (sequence from human genome, NG_008437.1, 469 pb). This promoter region is expected to drive expression specifically in neuronal cells.
This category contains vectors with the GUCY2Dmut transgene. They are available with a VSV envelope and a IN(WT) integrase.
GUCY2Dmut — Guanylate cyclase 2D
GUCY2Dmut cDNA encodes a mutant guanylate cyclase 2D with two substitutions: 837 D>E, 838 R>S (modified sequence from human genome, NM_000180.3, 3641 pb). This enzyme is guanylate cyclase 2D with dominant negative substitution D837E and R838S. These mutations have been associated in humans with cone dystrophies.
Transient expression of the transgene is achieved through modifications of the encapsidation plasmid, by making the vector integrase deficient. With these vectors, a broad range of transgenes and promoters are available.
IN(D64V) — HIV-1 integrase, catalytic site mutant (D64->V)
IN D64V is the HIV-1 integrase with a substitution in the catalytic site (64 D>V). This viral enzyme is normally responsible for the recombination of the vector genome with host cell chromosome. This activity is inactivated due to the mutation. Vectors produced with this integrase are deficient for integration.
IN(N) — HIV-1 integrase, basic region mutant (262 AAH->RRK)
IN N is the HIV-1 integrase with a substitution in the N basic region (262 AAH>RRK). This viral enzyme is normally responsible for the recombination of the vector genome with host cell chromosome. This activity is inactivated due to the mutation. Vectors produced with this integrase are deficient for integration.
IN(LQ) — HIV-1 integrase, basic region mutant (186 K>Q, 214 Q>L, 216 Q>L)
IN LQ is the HIV-1 integrase with a substitution in the LQ basic region (186 K>Q, 214 Q>L, 216 Q>L). This viral enzyme is normally responsible for the recombination of the vector genome with host cell chromosome. This activity is inactivated due to the mutation. Vectors produced with this integrase are deficient for integration.
