Transfection

into E.coli plasmid to amplify the vector, following by isolation of plasmids using specific Mini-prep Kit. Then, plasmids are transfected into Human Embryonic Kidney 293 cell line (HEK293 cell) or CH12F3 cell line. To determine the efficiency of transfection, we monitored green fluorescent protein (GFP) reporter construct expression by conducting either immunofluorescent assay or fluorescence-activated cell sorting (FACS) 14. We transduce the virus containing miR-125a vector into primary B cells that

Introduction For almost 10 milleniums, with “treatises as much as 2000 years old” giving advice on selective breeding, humans have carried out genetic manipulation on other organisms for their own benefit. Humans manipulate the genetics and genetic frequencies of both individual organisms and entire populations in order to raise the frequency of existing or transgenic traits we find useful or desirable. Human manipulation occurs for a large number of reasons, including; increasing profit/productivity

Alzheimer’s disease Dementia is a broad category of brain diseases that cause long term loss of the ability to think and reason clearly that is severe enough to affect a person’s daily functioning. Alzheimer’s disease (AD) is the most common form of dementia. It primarily affects mainly older adults. Memory impairment is an essential feature of AD. Human central nervous system (CNS) is composed of neuron and glial cell ( astrocytes, oligodendrocytes, microglia) which act as structural support of

Other vector variations are also available to provide added convenience, such as GFP for transfection monitoring or lenti-backbone for viral delivery. For researchers who perform mRNA microinjection or mRNA transfection. The applications for CRISPR Cas 9 are endless. There are two types of editing that are commonly used: homologous recombination and non-homologous end joining(NHEJ). Homologous recombination

Materials and methods The levels of microRNA-22-3p (miR-22-3p) and TP63 were analyzed by RT-qPCR and/or Western blot. Cell proliferation was measured using MTT assay. Cell invasion and migration were evaluated using Transwell assay chambers. Prediction of a regulatory relationship between microRNA-22-3p and 3'-UTR of TP63 mRNA was performed by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. OC cell lines Lung cancer cell lines H292, PC-9, CL1-5, A549 and normal NHBE

. Introduction HIV (Human Immunodeficiency Virus) is able to integrate its genome into the host cell genome during infection (Ebina, Misawa, Kanemura, Koyanagi, 2013). This can lead to AIDS (acquired immunodeficiency syndrome) if it is left untreated. AIDS is caused by a very efficient and sneaky retrovirus. HIV can remain dormant for years once its DNA is in the genome of the host cells. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats) are segments of prokaryotic

Transfection and selection was pursuant to previous description [40]. The empty vector served as the negative control. For the EGFP-LC3 cell line, the pEGFP-LC3 overexpression plasmid was previously prepared in our lab and used for cell transfection, followed by G418 (100 μg/ml) selection. Transient Transfection Cells were transfected with the miR34a, an antisense miR34a inhibitor (anti-miR34a), siSH3GL3

Accelerated Sensor of Action Potentials 1 The authors were trying to achieve a goal that they describe as a major goal in the field of neuroscience. They were looking for some method of reporting electrical activity in neuronal populations, specifically optical reporting. They developed a voltage sensor that they named Accelerated Sensor of Action Potentials 1 or ASAP1. This allows for GFP to be inserted into an extracellular loop of a voltage-sensing domain (VSD) and making fluorescence responsive

individual’s cells through the use of viral vectors, to replace the mutated ADA gene that is non-functioning or functioning incorrectly. In order for this process to be viable multiple techniques are used, including: restriction enzymes, ligation, PCR and transfection. Through these processes an individual is able to As there are 70 known mutations on the ADA gene currently that can cause ADA deficiency, I have chosen to focus on one specific mutation’s gene therapy. This mutation is a substitution mutation

HEK293A cells were used in experimentation and transfected with the ASAP1 plasmid. The HEK293A cells, which are very popular for transfection, were chosen as they are easy to reproduce, maintain, and have a high efficiency of transfection. HEK293A cells also have minimal processes that need to be accomplished in order to preform voltage-clamp experimentation. Transfection is simply the process by which