Strong and ubiquitous expression of the gene of interest
Single or double expression cassette with choice of reporter gene
Target gene expressed from CMV, EF1, or MSCV promoter
Choose from FIV- or HIV-based vectors
Gene expression from constitutive promoters The multiple cloning site (MCS) located downstream of a internal promoter allows convenient cloning of your gene of interest. Choose from three different promoters for expression of your gene--CMV (Cytomegalovirus), MSCV (Murine Stem Cell Virus), UbC (Ubiquitin C), PGK (Phosphoglycerate Kinase) or EF1 (Elongation Factor 1α). The table below describes the expression level and possible applications for each promoter.
Reliable delivery to dividing or non-dividing cells When used with lentiviral packaging plasmids, your cDNA constructs can be packaged into VSV-G pseudotyped viral particles and delivered into a wide range of mammalian cells with high efficiency. Stable cell lines can be generated that express the gene of interest. The system can also be used to overexpress genes in model organisms.
Convenient selection of transduced cells In addition to the pCD-MCS one-promoter vectors, which feature the Multiple Cloning Site (MCS) downstream of the CMV promoter, the pCD system offers the option of a second expression cassette downstream of the MCS to express the puromycin resistance gene or copGFP as a reporter under the control of a constitutive human elongation factor 1α (EF1) promoter, which is functional in most cell lines.
Efficient coexpression of target and reporter with T2A peptide SBI’s most recent additions to the cDNA vector collection confront the problems of promoter interference and imbalanced expression by incorporating a “self-cleaving” T2A peptide derived from the insect virus Thosea asigna to mediate coexpression of a reporter gene with the target cDNA. This allows an easy way to track cells actively expressing your gene of interest.
HIV-based lentivectors
Description
Vector Maps
The CD51X Dual Promoter Series High level of expression from either the CMV, UbC or MSCV promoter. The multiple cloning site (MCS) located downstream of a promoter allows for convenient cloning of your gene of interest. The SV40 polyadenylation signal in both vectors enables efficient termination of transcription, resulting in high levels of steady-state mRNA. Downstream of your expression cassette is an EF1 promoter driving the expression of either the Puro, GFP, RFP, Neo or Hygro marker genes in the Single marker options. Lentivector CD515B-1 MCS does not have a unique EcoRI site, as there is an additional EcoRI site in the Hygro marker gene.
SBI also offers a Dual marker cDNA expression vector that features both GFP and Puro markers that are co-expressed (cat# CD513B-1).
The CD52X EF1 Promoter Series with T2A Co-expressed markers SBI’s cDNA expression vectors using the EF1 promoter driving expression also incorporates the 2A-like sequence (T2A) from the insect virus Thosea asigna to mediate the co-expression of a reporter gene with the target cDNA. Reporter genes have been cloned at either the first or second positions relative to the T2A element, and both markers achieved high expression levels at either position.
The CD53X EF1 Promoter Series with IRES Co-expressed markers SBI’s cDNA expression vectors using the EF1 promoter driving expression also incorporates the Viral IRES (Internal Ribosomal Entry Site) sequence to mediate the co-expression of a reporter gene with the target cDNA. Reporter genes have been cloned at either the first or second positions relative to the IRES element, and both markers achieved high expression levels at either position.
MSCV Promoter Series The MSCV CpG-deficient promoter is the 5’-LTR promoter of murine stem cell virus that is incorporated into the 3’ HIV LTR of the cDNA expression lentivector. After integration into genomic DNA, the 3’MSCV/LTR will replace the 5’LTR and provide a high level of expression of the target gene and the downstream reporter gene. This promoter configuration has durable activity in most hematopoietic, embryonic and stem cells.
Clone-it Ligase free Cloning Series The CD520 series with the T2A configuration is also available in Clone-it ligase free cloning lentivectors (cat.# LF520A-1, LF520B-1, LF522A-1). The Ligase-free Cloning Site (LCS) replaces the Multiple Cloning Site (MCS) in the CD520A-1 type lentivectors. More Clone-it™ product information.
FIV-based lentivectors
Description
Vector Maps
—CMV promoter: promotes a high level of expression of your gene of interest in a wide variety of cell lines. —Multiple Cloning Site (MCS): for cloning the gene of interest in MCS located downstream of CMV promoter. —Hybrid CMV-5LTR promoter: provides a high level of expression of the full-length viral transcript in producer 293 cells.
Parikh H., Carlsson E., Chutkow W. A., Johansson L. E., Storgaard H., Poulsen P., Saxena R., Ladd C., Schulze P. C., Mazzini M. J., Jensen C. B., Krook A., Björnholm M., Tornqvist H., Zierath J. R., Ridderstråle M., Altshuler D., Lee R. T. , Vaag A.,1,2 Groop L. C., and Mootha V. K. TXNIP Regulates Peripheral Glucose Metabolism in Humans. Plos Med. 4 (5): e158, 2007. (PDF) »
