What is the WPRE sequence?
Woodchuck Hepatitis Virus (WHV) Posttranscriptional Regulatory Element (WPRE) is a DNA sequence that, when transcribed, creates a tertiary structure enhancing expression. The sequence is commonly used in molecular biology to increase expression of genes delivered by viral vectors.
What is WPRE in AAV?
WPRE is a 600 bp long tripartite element containing gamma, alpha, and beta elements, in the given order [16] and contributes to the strong expression of transgenes in AAV systems [11-13]. It also enhances the expression of a transgene lacking introns.
Is WPRE a promoter?
WPRE-mediated enhancement of gene expression is promoter and cell line specific.
What is PGK promoter?
The Pgk-1 promoter is in a region rich in nucleotides G and C. This promoter can efficiently drive high levels of expression of reporter genes such as E. coli lacZ and neo. We have determined that the 120 bp upstream of the transcription start site functions as a core promoter.
What is CBh promoter?
The CBA promoter displayed mostly ubiquitous and high neural expression, but substantially lower expression in motor neurons (MNs). We report the creation of a novel hybrid form of the CBA promoter (CBh) that provides robust long-term expression in all cells observed with CMV or CBA, including MNs.
What is an expression cassette in gene therapy?
From Wikipedia, the free encyclopedia. An expression cassette is a distinct component of vector DNA consisting of a gene and regulatory sequence to be expressed by a transfected cell. In each successful transformation, the expression cassette directs the cell’s machinery to make RNA and protein(s).
Is PGK a strong promoter?
It is interesting that PGK, which is a weak promoter in mammalian cells, can behave as a fairly strong promoter in fly cells. It is also interesting that mammalian promoters could be functionally conserved, at least to some extent, in such evolutionary divergent host cells.
Is ef1a a strong promoter?
The EF‐1α promoter is known as one of the strongest promoters in various mammalian cell lines 32, and the CAG promoter has been used frequently to drive strong gene expression in mammalian cells.
Does WPRE enhance the expression of transgenes in simple retrovirus-based genes?
WPRE enhances the expression of transgenes carried by MLV-based vectors. Since the WPRE is functional in settings as different as those of WHV and HIV-based vectors, it was considered likely that it would also be functional when incorporated into a vector derived from a simple retrovirus.
Do the RRE and WPRE control cytoplasmic genomic RNA abundance in lentiviral vectors?
The RRE and WPRE are required for optimal vector titre and transgene expression but do not control cytoplasmic genomic RNA abundance All lentiviral vectors derived from HIV-1 have the major splice donor (SD1) in the 5’ leader region and this sequence is part of the genomic RNA dimer structure 42.
Are viral sequences essential for transduction efficiency in lentiviral vector genomes?
Herein, we have conducted a systematic analysis of the viral sequences present in a lentiviral vector genome to determine which regions are not essential for transduction efficiency and could potentially be eliminated.
How well do mut6 WPRE vectors with WPRE function in vivo?
In summary, our data show that vectors with the mut6 WPRE adequately function in vivo for extended periods of time to ensure high levels of transgene expression and functional correction in a murine model of ex vivo gene therapy for WAS.