Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 921 results for: tro

Showing: 181 - 200 of 921 results

  1. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...produced in high-titer retroviral stocks generated by the transfer of a retrovirus into the packaging cell...are interested in using lentiviral vectors to introduce your favourite gene into your favourite cell line...CRISPR/Cas9 components, and fluorescent sensors. Retroviral vector background Until the development of viral...cells, labs use viral vectors and particularly retroviral vectors because they target a broader range of...into the genome is more efficient. The first retroviral vectors used were derived from the Moloney Murine...MoMLV, lentiviruses, a separate genus of the Retroviridae family including human immunodeficiency virus...infect non-dividing cells is not restricted to in vitro cell culture as lentivirus-derived vectors are capable...

  2. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...inefficiently our edits were being introduced. Although we were able to introduce the desired mutations in genes...The CRISPR/Cas9 com­plex is very efficient at introducing double stranded breaks (DSBs) into genomic DNA...generating gene knockouts, NHEJ repair does not allow introduction of specific sequence changes. To generate ...Accomplishing this usually involves simultaneously introducing a homologous DNA repair template, such as a sin­gle-stranded...in our lab. Our goal was to use the system to introduce disease-causing mutations in iPSCs (or correct... but had never been systematically analyzed: introducing silent CRISPR/Cas9 blocking mutations in either...target sequence once the desired edit has been introduced. We found that blocking mutations increased the...

  3. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...Callaway EM. Retrograde neuronal tracing with a deletion-mutant rabies virus. Retrograde neuronal tracing...a variety of cargoes, genetically engineered neurotropic viruses emerged as powerful tools for visualizing...that cell, and nascent RABV cross synapses to retrogradely infect neurons that project to the starter cell...occurs, subsequent cycles of viral replication and retrograde transmission occur, spreading the virus through...RABV G protein is necessary and sufficient for retrograde transmission (Etessami et al., 2000, Wickersham...2007). Thus, RABV transsynaptic spread can be controlled by selectively supplying the G protein in trans...released from the starter cell and transmits retrogradely to first order neurons, via entry mediated by...

  4. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...AAV9 serotypes have strong neuronal tropism, AAV5 targets both neurons and astrocytes, and AAV4 targets ...receptor subunits or neurotrophic factors. Another option uses optogentics to control neuronal activity,..., behavioral neuroscientists can now precisely control where and when certain genes are expressed to investigate...specific behavior. This blog post will give an introduction to behavioral neuroscience, and address experimental... in a laboratory, a high level of experimental control is required, because seemingly tiny external factors...Experimenter Interaction should be minimal and strictly controlled; optional handling animals Experiment Behavioral...channels that are chemically activated. Both are introduced in neurons and depending on the choice of channel...

  5. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    ...such as prototrophy, the ability to synthetize all compounds needed for growth, or auxotrophy, the inability...synthesize a compound needed to grow. Auxotrophy and prototrophy come into play during cloning if you’...lab created a yeast prototrophy kit containing 23 plasmids to complement auxotrophy. Negative selection...antibiotic resistance cassette and you aren’t introducing an additional antibiotic resistance cassette ...plasmid backbone contains your gene of interest. Auxotrophy Besides antibiotic selection, you can select ...you’ve designed your experiment using an auxotrophic strain of bacteria (one that doesn’t synthesize an ...amino acid), and a plasmid that complements the auxotrophy (i.e., synthesizes the critical amino acid). ...

  6. Technical Design of a Western Blot

    Type
    Blog Post
    ...expensive Nitrocellulose or low-fluorescence PVDF membranes Increased fragility (nitrocellulose) or increased...stains as loading controls: An alternative to high-abundance single protein controls in semi-quantitative...either DTT or β-mercaptoethanol (BME). DTT is a stronger reducing agent than BME and has a less… distinctive...life when stored properly. Because BME is not as strong as DTT, you’ll have to use different concentrations...Membranes The two membrane options are PVDF and nitrocellulose. PVDF is a sturdy membrane that can be stripped...membrane, it also has higher background noise than nitrocellulose membranes. Pro tip! You can visualize proteins...methanol and place it on a light box after drying. Nitrocellulose, on the other hand, doesn’t require any pre-wetting...

  7. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...Here's what makes these new backbones special: Strong expression: A CMV promoter drives robust gene expression... a test drive! We validated viral packaging and strong expression after inserting EGFP and provide step-by-step... Looking for more? Explore our empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing...Image courtesy of IPI.   Repurposing retrotransposons for DNA editing By Emily P. Bentley For RNA-guided...beyond Cas9 with STITCHR: a combined CRISPR / retrotransposon system with the pithy name "site-specific target-primed...insertion through targeted CRISPR homing of retroelements." The Abudayyeh-Gootenberg Lab developed this...this tool based on several new site-specific retrotransposon elements they identified and characterized ...

  8. New Viral Vectors - Winter 2025

    Type
    Blog Post
    ...viral prep pAAV-Ef1a-fDIO EYFP AAV5. AAV8, AAVrg Control Karl Deisseroth New serotype (multiple) rAAV-CAG...pAAV-AiE0452h-minBG-SYFP2-WPRE3-BGHpA (Alias: CN2237) AAV-PHPeB Control Jonathan Ting New viral prep AiP12787 - pAAV-...minBG-SYFP2-WPRE3-BGHpA (Alias: CN2787) AAV-PHPeB Control Jonathan Ting New viral prep AiP12610 - pAAV-...pAAV-AiE0780m-minBG-SYFP2-WPRE3-BGHpA (Alias: CN2610) AAV-PHPeB Control Jonathan Ting New viral prep AiP13755: pAAV-AiE0743m...10aa-H2B-WPRE3-BGHpA (Alias: CN4496) AAV-PHPeB Control Jonathan Ting New viral prep AiP14825: pAAV-AiE0441h...prep pAAV_BiSSTe10_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New viral prep pAAV_BiSSTe10_ChR2... pAAV_BiCHATe27_dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New viral prep pAAV_BiCHATe27_...

  9. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...concepts discussed in the paper. The transcriptional control of multiple loci is deftly coordinated by the eukaryotic...systematically delete/overexpress endogenous genes or to introduce heterologous genes, but the trend of research ...tinkering with regulatory networks and multiplex gene control. However, these approaches are often met with the...Modifying the CRISPRi system for multidirectional control The CRISPR interference (CRISPRi) model consists...CRISPRi system for more flexible multidirectional control, a new study repurposes the sgRNA to include modular...The use of regulators allows a greater degree of control of desired expression levels dependent upon the...small allowing multiple sgRNA constructs to be introduced simultaneously in a single plasmid. The functionality...

  10. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...is sufficient. Controls: Controls can make or break an experiment. The best controls for a colony PCR... They also serve as controls for your PCR reaction. Running a no template control PCR reaction for detecting...bacterial debris. Use a positive control. A good positive control is bacteria transformed with the same...primer design. Use a negative control strain. A good negative control strain is an untransformed culture...type of control is especially important for insert-specific primers. If your negative control amplifies...number of background colonies on your no ligation control plate. The more background, the more colonies you...backbone vector with and without an insert. These controls are quick references you can use when you run ...

  11. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...basics: What is optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford...1984; Harz and Hegemann, 1991). These proteins control the flow of electric charges across cell membranes...opsin genes in targeted cells makes it possible to control neuronal activity with a specificity far greater...can be achieved using pharmacological or electrophysiological methods (Deisseroth, 2011; Mei and Zhang... to flow across the axon’s membrane, thereby controlling neural activity (Bernstein and Boyden, 2011; ...usually drive weak expression of downstream genes. Strong cell-specific promoters, on the other side, are...; Hudry and Vandenberghe, 2019). How to achieve strong opsin expression in specific neuronal cell types...

  12. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...Transposable elements (TEs): Retrotransposons  Class I TEs are also known as retrotransposons. They transpose by...those without (non-LTR TEs). LTR retrotransposons are like retroviruses both in their structure and mechanism...which are required for LTR retrotransposon transposition. Non-LTR retrotransposons contain two open reading... Figure 1: Overview of retrotransposon transposition. Retrotransposons are mobilized by a ‘copy-paste...such as the ~11 kB cDNA for dystrophin, the gene mutated in muscular dystrophy, require truncation to fit...of transposition: Class I TEs, also called retrotransposons, and Class II TEs, also called DNA transposons...into new target sites. This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode...

  13. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...probability of artifacts. Off-target controls A seldom used control for targeted CRISPR experiments is ... how to predict and minimize them, and ways to control for and quantify them when they can’t be avoided... system is an RNA-guided nuclease that either introduces a ssDNA nick, a dsDNA break, or is engineered...placement (e.g., mutating a specific amino acid or introducing a specified edit). However, if your design does...putative off-targets would be suitable. How to control for off-target events Sometimes there just isn’...applications, this is ok, but it still must be controlled for and acknowledged as a potential limitation...to perform key experiments on is a good way to control for off targets as well as clonal differences. ...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...sample. Troubleshooting Tips   Figure 3. Summary of plasmid multimerization troubleshooting tips...capillary gel electrophoresis. BioProcess Int. 2006 Sep;4(8):38-40. Summers D. Timing, self-control and a sense...sometimes observe something odd in our uncut DNA control: a band or two appear on an agarose gel at notably...agarose gel Running undigested DNA on a gel as a control when performing a diagnostic digest is a simple...sequencing of our plasmid samples during our quality control process. Since MiSeq involves sequence assembly...identification of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique that can detect... detect multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene...

  15. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...Noninvasive Controlling Stimulation ExogenousEasily controlled EndogenousNot easily controlled Unique...spatiotemporal control of a small subset of neurons is necessary, use optogenetics. If broad control of whole...boundaries in neuroscience by granting targeted control over neuronal activity. While they serve similar...optogenetics differ are: Timing Targeted manipulation Controlling stimulation Invasiveness  Choosing the best ...Chemogenetics does not offer the same level of control over timing. Because chemogenetics relies on chemical... can reach target cells throughout the body. Controlling Stimulation Some studies rely on delivering reversible...stimulation to receptors. Optogenetics offers better control over these factors as the light source can be easily...

  16. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ... or chaotropic salts. Capillary electrophoresis Quantifying DNA using capillary electrophoresis is ... using gel electrophoresis. Except, it’s smaller. And automated. Unlike gel electrophoresis, you only ...wavelength that they maximally absorb. The spectrophotometer measures these absorbances using UV-transparent...absorb best at 280 nm and organic compounds and chaotropic salts maximally absorb at 230 nm. The A260/A280...specific for double strand DNA (as compared to spectrophotometric methods that measure all nucleic acids). ... sample concentration for you. Agarose gel electrophoresis Though not the quickest way to quantify DNA...methods can’t tell you and you don’t need a spectrophotometer or a fluorometer to quantify DNA this way....

  17. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ...have learned how to introduce genetic materials into cells in the lab. The introduction of foreign DNA or...research. There are multiple ways foreign DNA can be introduced into cells including transformation, transduction...plasmid of interest. Scientists can also use electroporation, the application of an electrical charge to...Transduction occurs when foreign DNA or RNA is introduced into bacterial or eukaryotic cells via a virus...Transduction is a common tool used by scientists to introduce different DNA sequences of interest into a bacterial...transduction. Scientists also use transduction to introduce foreign DNA into eukaryotic cells, like mammalian... process by which foreign DNA is deliberately introduced into a eukaryotic cell through non-viral methods...

  18. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...Finsterbusch, Michaela, et al. "Patrolling monocytes promote intravascular neutrophil activation and glomerular...same laser) cooperate to cause a higher-energy electronic transition in a fluorescent molecule (see figure...epifluorescence microscope and GFP fusion proteins controlled by promoters specific to the immune lineage, ...movements in real time. This technique was first Introduced to immunology by three papers in the journal ...of pathogen-immune cell interaction relied on in vitro studies where it was difficult to apprehend the ... The video above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph...infection with S. aureus (red). You can then see neutrophils swarming around S. aureus and eating the bacteria...

  19. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

    Type
    Blog Post
    ...pouring in! While our lentiviral service is going strong, the AAV service has shown incredible growth this...EBFP-Cre Available in serotypes AAV1, AAV5, and AAV retrograde (the most popular form), this viral vector is...deliver DNA encoding Cre recombinase under the control of the synapsin promoter. It can be used to drive...genes to various brain regions. You can partially control which cells are successfully infected by an AAV...serotype, but occasionally you may want to further control where your gene of interest is expressed.  A cell-type...cell-type specific Cre can provide you with that control in a modular way. For example, say you would like...contains a Cre-dependent mCherry cassette under the control of an otherwise ubiquitous promoter. You could ...

  20. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ... for introducing one modification into the mouse genome, they are not commonly used to introduce multiple...opposed to ordering control mice from a stock center. This serves as a better control since the experimental...series, we covered techniques that can be used to introduce genetic modifications into mouse embryos. But ...knockout lines and Cre-lox lines, and how to properly control for genetically engineered mice in your experiment...mutations. This is because as more mutations are introduced into a single embryo, the likelihood that a mouse...genetic engineering should be used to separately introduce each mutation independently into different mice...not a waste. Best practice is to use littermate controls in your experiment - use mice that were bred in...
Showing: 181 - 200 of 921 results