Detail Introduction: Duchenne muscular dystrophy (DMD), caused by out-of-frame mutations of dystrophin gene, is a progressive degenerative disease resulting in cardiac and smooth muscle dysfunction in approximately 250–300,000 boys and young men worldwide. Previous studies showed that AAV-delivered CRISPR gene editing tools can reframe the mutated dystrophin gene and restore dystrophin expression in vitro and […]
Detail Introduction: Around one in 1000 newborns are suffered from congenital disabling hearing loss, among which, mutations in OTOF gene (encoding otoferlin protein) accounting for 2.3–10% of patients, and over one thousand pathogenic mutations have been identified within this gene. Therefore, a postnatal supplement of otoferlin cDNA into the inner ear might be a promising […]
Detail Introduction: Duchenne muscular dystrophy (DMD), owing to the gene mutation of dystrophin, is a severe genetic disease resulting in body-wide muscle degeneration and necrosis in boys and young men. Lots of evidence demonstrates that an important early pathogenic event from DMD patients is the abnormal elevation of intracellular Ca2+, resulting from the dysfunction of […]
Adenovirus protocol Download For most serotypes, adenovirus infection is mediated by the high-affinity binding of the fiber-knob region to a receptor of target cell, named as the coxsackie-Ad receptor (CAR) [17]. Upon attachment, interaction between the penton-base Arg-Gly-Asp (RGD) and cellular αv integrins, which can stimulate actin polymerization, leads to internalization of the virus into […]
Adenovirus protocol Download Adenovirus is a non-enveloped, 90-100 nm diameter virus presenting icosahedral symmetry (Figure1A). Human adenovirus, containing a linear, double-stranded DNA genome, which is approximately 36kb, is wrapped in a histone-like protein and has two inverted terminal repeats (ITRs) of 50-200 bp, which act as origins of replication, accompanied by a complex series of […]
Adenovirus protocol Download a) Advantages of Adenovirus-mediated gene delivery Adenovirus has been developed into a preferred candidate for creating viral vectors for gene therapy due to various advantages.1) Well tolerated, with post-infection viability of the host cells being almost 100%.2) Great packaging capacity (up to 8kb).3) Broad range of infectivity. Adenovirus can infect both dividing […]
Adenovirus protocol Download Adenovirus has been proved as an excellent gene therapy vector. To date, more than 535 clinical trials have been carried out using adenovirus vectors for gene delivery [7], and promising gene therapy outcomes from recombinant adenovirus have been achieved from clinical trials for a great number of diseases (Table 1), especially for […]
Adenovirus protocol Download Adenovirus (AdV) is a member of the family Adenoviridae, whose name derives from their initial isolation from human adenoids in 1953 [1]. It is a medium-sized (90-100nm) and non-enveloped virus with an icosahedral nucleocapsid containing a double-stranded DNA genome. With a broad range of vertebrate hosts, 100 serotypes have been isolated, and […]
AAV protocol Download AAV Production Protocol–Genemedi 1. AAV plasmid construction The gene of interest is cloned into one of the ITR/MCS-containing AAV vectors to generate AAV-GOI. The purity and RNAse contaminants of AAV viral plasmid should be taken into consideration. 2. AAV packaging The recombinant AAV viral plasmid AAV-GOI is co-transfected into the AAV-293 cells […]
AAV protocol Download Adeno Associated Virus (AAV) For normal tissues or organs, such as heart, liver, kidney, breast, pancreas, ovary, brain, eye, skeleton muscle, adipose tissue, etc., Genemedi systematically organizes the corresponding optimal AAV serotype, gene delivery method and injection volume for mouse and rat tissue infection. Recommended AAV gene delivery methods for different organs is […]
