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... AAV vectors expressing RGB proteins under the control of an inducer, i.e. tet-off transactivator (tTA...Subscribe to Viral Vector Blog Posts Additional Resources on Addgene.org Search for AAV Vectors to use with...   Adeno-associated viral (AAV) vectors are the most frequently used gene-transfer tools in the study...infect the PNS or the CNS much better than the AAV vectors they were derived from. Additionally, AAV.PHP.eB... of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids were packed...AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for?  1. Multi-color...multi-color labeling is to inject a mix of AAV vectors that constitutively express a red, green, or blue...

...posts related to viral vector protocols and tips Read all of our viral vectors blog posts Resources on...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ...titrating AAV by qPCR, viral DNA is amplified and monitored in real time. When the PCR is complete, the results...issue with qPCR is that the results can vary by a factor of 2. This means that if you set up two identical.... This partitioning reduces the number of PCR inhibitors per reaction and allows for enhanced detection...up with poor droplet quality. Using a droplet generator, the sample and the droplet generation oil are... = Volume of virus in reaction mix D = Dilution factor of virus Most droplet readers have a few channels...

... plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. Today, it'... in this post:  New compact, efficient prime editors A versatile and precise system to improve on-target...proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old antibody...antibody new tricks   New compact, efficient prime editors By Susanna Stroik The Liu lab recently used phage-assisted...protein engineering to generate seven new prime editors, PE6a–g, that overcome some current prime limitations... and PE6b are 500–800 bp smaller than previous editors, but just as effective and with no additional off-target...protein engineering yield compact, efficient prime editors. Cell, 186(18), 3983–4002.e26. doi: 10.1016/j.cell...

...the activator protein cannot bind. After an inducer binds to the activator protein, the activator protein...In this system, the activator rtTA (reverse tetracycline-controlled transactivator) is normally inactive...the binding of RNA polymerase and transcription factors. Since the promoter region drives transcription...agents, temperature, and light are all examples of factors that can lead to the induction of a promoter. Below...inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is another popular...protein purification. When arabinose is absent, regulatory protein AraC binds O and I1 sites upstream of...addition to arabinose, cAMP complexed with cAMP activator protein (CAP) can also stimulate AraC binding ...

...zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen total variants deposited... plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles. Here's what...systems for specific applications! Delivering prime editors (PEs) via engineered virus-like particles (eVLPs... contain no genetic material, which ensures the editor will only be transiently present in transduced ...site-specific serine recombinase. First, the prime editor installs the recombinase “landing sites,” then ...landing sites on opposite DNA strands. When the prime editor releases the DNA, the two 3’ flaps containing the...-like particles for transient delivery of prime editor ribonucleoprotein complexes in vivo. Nature Biotechnology...

...G-protein-coated RABV infects cells via the G receptor. In neurons, the G receptor is restricted to presynaptic nerve...Control Gene Expression with FLEx Vectors Resources on Addgene.org AAV Vector for TVA Expression AAV for Rabies...housed in specific locations and give rise to progenitors during growth and development or that blood flows...to visualize how neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability to efficiently...through their interactions with target cell surface receptors (Callaway 2008). Based on this, cell type specificity...EnvA-pseudotyped RABV, and only expressing the EnvA receptor (TVA) in user-defined cells. For example, a researcher... via entry mediated by the G protein/G-protein receptor. Once in first order neurons, the G-deleted RABV...

...acquire during their PhD and postdoctoral training. Addgene executive director Joanne Kamens has written ...were many changes to the  Board of Directors and the Executive Director changed twice. I had to learn how... to report to the Board of Directors and liaise with the Executive Director. As members changed, I had...management-for-scientists-managing-vs-leading. The National Postdoctoral Association (NPA) outlines 6 Core Competencies... and Management skills”. The NPA states that Postdoctoral scholars should have the skills and techniques... levels. These skills will also help the person mentor others more successfully. But how does one gain... leadership experience). I was involved with postdoctoral issues first at Brigham and Women’s Hospital...

...the vector backbone you cloned your insert into and fill out the appropriate boxes with the vector backbone...backbone manufacturer, vector backbone size, total size of your vector + insert, any modifications you ...standard conditions (37 °C, LB, antibiotics, etc.) and stored as glycerol stocks We will need information on...provide any full or partial plasmid sequences and any vector maps. The more sequence data available, the better... was a part of the insert (when cloned into the vector backbone). If your plasmid has more than one insert...you made to the backbone, and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer...enter "Feng Zhang (Addgene plasmid #52961)".   The vector type should describe how your plasmid is intended...

... — but critical — factor for successful experiments or therapies. Using AAV vectors with enhancers that... plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. Here's what...what you'll find in this post: Glutamate indicators with increased sensitivity and tailored deactivation...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation...abundant neurotransmitters, but it is challenging to monitor because only a small number of glutamate molecules...and they are rapidly taken up by postsynaptic receptors and transporters. The intensity-based Glutamate-Sensing...fusions for membrane display, they recommend the NGR vectors for most experiments. These new tools promise to...

...point will be those that have excised the vector backbone. Vectors pTOX4, pTOX5, and pTOX6 express the YhaV...Find the allelic exchange vectors here! ShadowR, a novel FLIM-FRET acceptor Figure 4: ShadowR is...levels. Chromoproteins in nature Compared to the history of fluorescent proteins in molecular biology research...research, chromoproteins have a shorter history. In 1987, scientists described an unusual blue-colored... selection markers for allelic exchange. These vectors contain a toxin gene (regulated by a rhamnose-inducible...leads to the desired mutation and excision of the vector backbone. To do this, rhamnose is added which induces...respectively, and all encode the amilCP chromoprotein. Vectors pTOX7, pTOX8, and pTOX9 express the YhaV, MqsR,...