Chemogenetics Plasmids
Type
Collection
...specific small molecules, allowing a scientist to control signaling within cells expressing these engineered...
We narrowed to 922 results for: TRO
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Addgene Packaged on Request: Scope of Service
TypeCollection...for the AAV1, AAV5, AAV6, AAV8, AAV11, or AAV Retrograde serotypes and 2 x 10 12 GC/mL for the AAV2 serotype... -
Antibody Plasmid Collection
TypeCollection...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs... -
CRISPR Plasmids - Prime Edit
TypeCollection...enzymes and related tools are available: PE2 - introduction of five mutations in RT enzyme PE3 - PE2 plus... -
Promega Plasmid Collection
TypeCollection...HaloTag (Link opens in a new window) technology puts control of the function and properties of a fusion protein... -
Kazuhiro Oka Lentiviral Vectors
TypeCollection...vectors to express your gene of interest under the control of a variety of promoters. Professor Oka and the... -
TALEN Guide
TypeCollection...new technology, and we hope to continue to have a strong scientific partnership with both the labs that ... -
Arf GTPase Family
TypeCollection...traffic, they also play critical roles at cilia, centrosomes, the plasma membrane, lipid droplets, the nucleus... -
COVID-19 Resources
TypeCollection... Plasmid Description Industry PI Due to export control regulations, MERS and SARS-CoV materials are available... -
CRISPR Plasmids - Tagging
TypeCollection...Yannick Doyon's lab deposited a plasmid which introduces an N- or C- terminal affinity tag (3xFLAG-2xSTREP... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...specificity, or retrograde transport in neurons. (b) Genetic cargo can be customized to control transgene expression... Capsid Published Tropism Model Organism Mouse Strain-Specific Tropism? Reference AAV-...and astrocytes Mice No (LY6C1) Hanlon et al., 2019 AAV-BI-28 CNS: neurons, astrocytes, oligodendrocytes...-Specific Tropism?" column. For a complete list of mice strains with LY6A-dependent tropism, see S3 table...derived from AAV9 using iTransduce, shows tropism toward astrocytes and neurons, with greatly increased efficacy... appears to show a broader tropism with a potential bias toward astrocytes (Chuapoco et al., 2023). Additionally...systemic AAVs with endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications... -
Using AAV for Neuronal Tracing
TypeBlog Post...developed a new AAV variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV... AAV2-retro vectors with a variety of transgene/promoter combinations. Check out our retrograde AAV blog...requires both antero- and retrograde labelling, synaptic restriction as well as controlled monosynaptic and polysynaptic...Selective retrograde transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal...bodies to the terminals of their axons; while retrograde tracing outlines neurons in the opposite direction...axons to their cell bodies. Anterograde and retrograde tracing take advantage of existing transport ...myosin, and enzymes for transmitter synthesis. Retrograde transport is used for transport of endocytosed... -
Plasmids 101: The protein expression toolbox
TypeBlog Post... What kind of control? On, off, tunable, and/or inducible? How long do you need control? Will temporary...temporary control (a few days) be enough, or do you want a permanent control mechanism? Where is the control...transcription instead of introducing a DNA double strand break. Transient introduction of CRISPRi or inducible...vector! On/off control all the time If your experimental system requires tunable control - or maybe you...price of tunable control These systems can be easily implemented for exogenously introduced proteins, as ...prior to introduction of the construct into cells. However, if you would like tunable control of an endogenously...forever? You don’t need to resort to the force to control protein expression in mammalian cells: easy to ... -
Viral Vectors 101: Types of viruses
TypeBlog Post...viruses: gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus. Gamma-retrovirus Retroviruses...type of gamma-retrovirus is Murine Leukemia virus (MLV). Naturally isolated gamma-retroviruses have been ...Lentivirus is a type of retrovirus and shares many common features with gamma-retrovirus including genome architecture... nucleus as gamma-retroviruses do. Fun fact: Since lentivirus and gamma-retrovirus are so closely related...expression (Robinson, 1982). To add insult to injury, retroviruses generally have moderate to high immunogenicity...immune response in the host. So why use a gamma-retrovirus in the lab? Well, they are essentially evolutionarily... types with a high efficiency. However, gamma-retrovirus infection is restricted to dividing cells only... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog Post...A common means to achieve ligand control is to fuse a mutant estrogen receptor (ER) ligand binding domain...PSD95-mcherry) under the control of Cre recombinase. After in-utero electroporation, this plasmid mix results...tracking for instance) can be controlled with FLP-FRT. For further temporal control, Tamoxifen inducible Cre...system, but it only gives you a single level of control without modification - either Cre is there or it...for site specific (and often cell type specific) control of DNA recombination and gene expression can be...addition, a variety of means to spatiotemporally control FLP and Cre expression have been developed. Read...FLP and Cre enable additional levels of genetic control. What is FLP FRT and how does it work? FLP recombinase... -
Antibodies 101: Flow Cytometry
TypeBlog Post...Include proper controls To ensure accurate data analysis, include a complete set of controls. Ideally, every...will have a negative control that does not express the target and a positive control that does. The negative...of experiments easier and faster to perform. Introduction to flow cytometry Flow cytometry allows users... (3): An optics system A fluidics system An electronics system Cells are first resuspended in a pressurized...fluorophore or fluorochrome-stained protein will emit a stronger photon than a cell expressing low levels of the... further the shift of a cell from the negative control, the higher the level of expression. A routine ...triton-x, saponin, or tween. Take care when using strong detergents like triton-x and tween as extended ... -
Important Considerations When Using AAVs
TypeBlog Post...larger amount of vector using a serotype with strong CNS tropism (4). If axonal transport is not desirable...receptor physiology and the effects of metabotropic and ionotropic influences on overall cell function. ... thanks to our guest blogger Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student at ...post was contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University...known is that different serotypes have different tropisms but, as you can see, AAVs of each serotype can... asking questions! Table 1: AAV Serotypes and Tropism Experiment Detection Method CNS AAV1, AAV2...>AAV8>AAV6>AAV5>AAV1>AAV2 This preferential tropism for different brain regions is largely due the ... -
Delivery Methods for Generating iPSCs
TypeBlog Post...Moloney murine leukemia virus (MMLV)-derived retroviruses. Retroviral vectors, such as pMXs or pMSCV, infect...delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic vector or multiple...10 kb) than other retroviral vectors. This makes it possible to use a polycistronic cassette that expresses...Mechanism Efficiency Safety Pros Cons MMLV-derived Retrovirus Integrating High Low Efficient and Stable Genome...Need to sequence cells to verify excision didn’t introduce mutations Adenovirus Non-integrating Low Medium...required for translational use. Viral Vectors Retroviral: Some of the first iPSCs were generated with ... corresponding endogenous pluripotency genes. Retroviral vectors are prone to incomplete silencing of ... -
Plasmids 101: Stringent Regulation of Replication
TypeBlog Post...DNA. Plasmids are said to be under stringent control when they depend on the host cell’s initiation ...replication independently are said to be under relaxed control, as they only require the host's replication machinery...replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...round of replication. Initiation is the primary control point of DNA replication in bacteria, so the process... the bacterial chromosome and for stringently controlled plasmids. Binding of IHF (green) redistributes...binding is necessary to initiate replication, controlling the amount of active DnaA available is one of...source, ATP binding is one more way for the cell to control DNA replication. A variety of other proteins also... -
Plasmids 101: The Promoter Region – Let's Go!
TypeBlog Post...target genes. The sequence of the promoter region controls the binding of the RNA polymerase and transcription...promoters is commonly employed in lentiviral and retroviral constructs and we will discuss these in a future... all the time while others are more carefully controlled. Regulated promoters might act only in certain... light. In the cell, promoters themselves are controlled by still other regulatory factors: enhancers,...Additional considerations CMV General expression mRNA Strong mammalian expression promoter from the human cytomegalovirus...some cell types. EF1a General expression mRNA Strong mammalian expression from human elongation factor...promoter hybrids. CAG General expression mRNA Strong hybrid mammalian promoter Constitutive Contains...