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 […]
CRISPR/Cas9 protocol Download Gene editing, also called genome editing, is a group of technologies to change the sequence of DNA in the genome. Several approaches to genome editing have been developed, including Zinc Finger, TALEN, and CRISPR/Cas9. Compared with the other gene editing tools, CRISPR/Cas9 system is faster, cheaper, more accurate, and more efficient, showing […]
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 […]
Lentivirus Protocol Download Lentivirus Production Protocol–Genemedi 1. Lentivirus plasmid construction The gene of interest is cloned into one of the LTR/MCS-containing lentivirus vectors to generate pLV-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration. 2. Lentivirus packaging The recombinant lentivirus viral plasmid pLV-GOI is co-transfected into the 293T with envelope […]
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 […]
Lentivirus Protocol Download This protocol is for the stable cell line construction based on puromycin selection. Day 1: Seed target cells in 24-well plates. The number of seeding cells differs according to the cell proliferation rate.Day 2: Target cells should be approximately 50%-70% confluent. For polybrene accessible cells, mix the culture medium with proper concentrations of polybrene. […]
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 […]
