What is CK8 promoter?
Thus, the CK8 promoter (MHCK7 with two copies of the MCK enhancer instead of the α-myosin heavy-chain enhancer) was used for intramuscular AAV8-mediated delivery of the growth hormone gene [53]. In mice treated with this construct, body length and weight were significantly higher compared to those in untreated mice.
What kind of promoter is needed to design skeletal muscle specific gene expression of a transgene?
The Des promoter was chosen since it is known to confer relatively high levels of skeletal muscle and heart-specific transgene expression35. The constructs were packaged using AAV serotype 9 (AAV9) to maximize skeletal muscle and cardiac-specific gene transfer36,37,38.
What is rh74?
Researchers in the Center for Gene Therapy will also soon launch a new trial in LGMD using another AAV vector, rh74, a virus isolated from rhesus macaque monkeys. This adeno-associated virus was developed at Nationwide Children’s and could address another key challenge to gene therapy: immune response.
What is micro-dystrophin?
micro-dystrophin is a gene therapy candidate that is designed to deliver the microdystrophin-encoding gene directly to the muscle tissue for the targeted production of the microdystrophin protein. It uses a harmless modified virus (AAVrh74) that has a high affinity for muscle tissue, allowing for targeted delivery.
What is MHCK7?
MHCK7 promoter (770 bp) is a hybrid promoter consisting of enhancer/promoter regions of murine muscle creatine kinase (CK) and alpha-myosin heavy-chain genes.
What is a regulatory cassette?
An expression cassette is a distinct component of vector DNA consisting of a gene and regulatory sequence to be expressed by a transfected cell. An expression cassette is composed of one or more genes and the sequences controlling their expression.
How do you identify a promoter sequence?
To find the promoter region, use Map Viewer to locate the gene within a chromosomal context. Then increase the value of the coordinates that surround the gene to a larger sequence that includes the promoter.
What is the promoter sequence in DNA?
A promoter is a region of DNA where RNA polymerase begins to transcribe a gene. Normally, promoter sequences are typically located directly upstream or at the 5′ end of the transcription initiation site (Lin et al., 2018).
What is micro dystrophin?
What do regulatory sequences do?
A regulatory sequence is a segment of a nucleic acid molecule which is capable of increasing or decreasing the expression of specific genes within an organism. Regulation of gene expression is an essential feature of all living organisms and viruses.
What are CRISPR cassettes?
Abstract. CRISPR/Cas, bacterial and archaeal systems of interference with foreign genetic elements such as viruses or plasmids, consist of DNA loci called CRISPR cassettes (a set of variable spacers regularly separated by palindromic repeats) and associated cas genes.
What is the function of CK8 promoter?
Thus, the CK8 promoter (MHCK7 with two copies of the MCK enhancer instead of the α-myosin heavy-chain enhancer) was used for intramuscular AAV8-mediated delivery of the growth hormone gene [53]. In mice treated with this construct, body length and weight were significantly higher compared to those in untreated mice.
What is the significance of ck8e 436?
The 436-bp CK8e construct carried a number of deletions in the enhancer and proximal promoter of muscle-type creatine kinase (Fig. 3) and was more active than the CMV promoter in a differentiated human myoblast culture [54]. Deletion of the poorly conserved regions in the promoter reduced its length and simultaneously increased its activity [31].
What is CK6 made of?
Thus, the construct CK6, consisting of an enhancer (206 bp) and a proximal promoter (358 bp) (Fig. 3), ensured high muscle specificity. However, the expression level in the muscles was ~12% compared to that attained using a similar construct with the CMV promoter; the expression level in the heart remained low [50]. Open in a separate window Fig. 3
What is the role of constitutive promoters in gene expression?
Many constitutive promoters are used to drive ectopic gene expression in various in vitro and in vivo contexts.