What is EF1 alpha promoter?
Human elongation factor-1 alpha (EF-1 alpha) is a constitutive promoter of human origin that can be used to drive ectopic gene expression in various in vitro and in vivo contexts.
What is SFFV promoter?
SFFV promoter provides the best level of transgene expression in Raji cells and LCLs. Since CMV and EF1α promoters allowed for ≤40.25% transgene expression in Raji cells further experiments were conducted using the SFFV promoter.
What is a U6 promoter?
U6 is a type III RNA polymerase III promoter commonly used for driving small hairpin RNA (shRNA) expression in vector-based RNAi. In the design and construction of viral vectors, multiple transcription units may be arranged in close proximity in a space-limited vector.
What are episomal vectors?
The Episomal iPSC Reprogramming Vectors are a non-integrating system that reprograms somatic cells into induced pluripotent stem cells (iPSCs). This product is a mixture of three vectors designed to provide the optimal system for generating transgene-free and virus-free iPSCs in a feeder-free environment.
What is Wpre sequence?
Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) is a DNA sequence that, when transcribed creates a tertiary structure enhancing expression. Commonly used in molecular biology to increase expression of genes delivered by viral vectors.
How strong is U6 promoter?
U6 promoter is constitutively active in a variety of cell types and maintains relatively high activity by providing approximately 4×105 transcripts per cell (29).
Why is U6 promoter used for shRNA?
U6 is the most optimal promoter to produce shRNAs, while CMV is used to express whole genes. Pol III genes and promoters do not get polyadenylated and thus are ideal for making small nuclear RNAs such as shRNA hairpins.
What is Episomal expression?
Integrating vector systems can deliver genetic material to a target cell with high efficiency enabling long-term expression of an encoded transgene. Episomal vector systems have the potential to avoid these undesired side effects, since they behave as separate extrachromosomal elements in the nucleus of a target cell.
What is the function of Episomes?
episome, in bacteria, one of a group of extrachromosomal genetic elements called plasmids, consisting of deoxyribonucleic acid (DNA) and capable of conferring a selective advantage upon the bacteria in which they occur.
Is WPRE expressed?
The increased expression from WPRE in LV vectors was not due to altered viral mRNA half-life or the rate of transcription; they therefore, concluded that WPRE probably acts post-transcriptionally. Our results support their findings and those of others that WPRE can increase transgene expression from RV and LV vectors.
Is WPRE translated?
The transcriptional activity of the WPRE was undetectable in the context of the lentiviral vector but the element was capable of translating a polypeptide. The WPRE was required to express high levels of the transgene and for that, the native form or mutated derivatives functioned equivalently.
Is U6 promoter inducible?
The most common inducible expression systems are based on tetracycline-regulated RNA polymerase III promoters. For all cell lines tested, we find that several inducible U6 and H1 promoters containing single tetracycline operator (tetO) sequences show high-transcriptional background in the non-induced state.
Where does the EF-1 alpha promoter come from?
The EF-1 alpha promoter, which offers a broad host range, is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1 (Uetsuki et al. 1989; Wan Kim et al.
How does the EF-1 alpha gene enhance gene expression?
We inserted the first intron of the human EF-1 alpha gene behind the MCMV and the HCMV promoter to enhance the gene expression through increasing RNA transcription and/or RNA stability.
How are EF-1 alpha promoters used in lentiviral vector systems?
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells. cPPT Element.
Is the EF1 promoter ubiquitous in PDRIVE plasmid?
EF1 promoter (Ubiquitous) in pDRIVE expresssion plasmid. The EF-1 alpha gene encoding elongation factor-1 alpha is an enzyme which catalyzes the GTP-dependent binding of aminoacyl-tRNA to ribosomes. EF-1α is one of the most abundant proteins in eukaryotic cells and is expressed in almost all kinds of mammalian cells.