...future) applications of lentiviral vectors! Lentiviral vector history and components In the early 1990s...design a viral vector system that could do both, Addgene depositor Didier Trono and collaborators turned to...Lentiviral vectors are one of the most popular and useful viral vectors in the lab. Advantages of lentivirus...dividing and non-dividing cells. Lentiviral vectors are the vector of choice for many CRISPR applications,...viral vector systems based on retroviruses like Moloney murine leukemia virus (MMLV). The vectors could...adeno-associated viral vectors can also target non-dividing cells, lentiviral vectors can carry much more...Lentiviral Plasmid FAQs. Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the lab...

...Genome Engineering Using FLEx AAV Vectors? Learn about Viral Vector Elements Read Our Primer on Optogenetics...improvement of optogenetic adeno-associated virus (AAV) vectors. Listed below are a few additional uses for FLEx...and Ghyselinck NB. A directional strategy for monitoring Cre-mediated recombination at the cellular level...PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed Boyden Bernardo Sabatini Wenbiao Chen Liquin...

...lentiviral vectors, you need an envelope protein on the virus’s surface and a corresponding receptor in the...lentiviral vector will infect, pseudotyping can help. In pseudotyping, you produce viral vectors using viral...glycoprotein. Lentiviral vectors are also great at infecting neurons and cause minimal inflammatory response, but...Sandoval S (2017) Pseudotyped Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods ...the host cell. Some of these envelope-receptor pairings are broad, allowing delivery into many cell types...virus. How to pseudotype To pseudotype your viral vector, you use the same protocol for generating lentivirus...also be cell type dependent: if a cell has few receptors for a specific envelope protein, it takes up less...

...method for targeting receptor expression as a ligand and receptor pair. Targeting Receptor Expression Using... translation to human patients. Picking receptors The receptor you choose is largely dependent on what... designer receptors exclusively activated by designer drugs, are G protein-coupled receptors (GPCRs), ...derived from muscarinic receptors. The most popular agonist for muscarinic receptor derived DREADDs has been...pharmacologically selective actuator modules, were developed with the goal of creating receptors with less endogenous...on the Addgene blog New to viral vectors? Browse our Viral Vectors 101 blog posts Read blog posts about... relies on genetically introducing engineered receptors that are only activated by otherwise inert drugs...

...specifically recognized by another factor, the receptor. The receptor could be something like an antibody...for multiplexed imaging for advanced monitoring of different factors, in different or the same cellular ...wide-range of uses to monitor the fundamental cellular building blocks. Physiological factors Unlike the previously...engineered into viral vectors, so if you want to use them, you’ll need to use the vectors (or redesign the ...directly proportionate to the amount of a given factor within a biological system. Basically, biosensors...antibody, enzyme, DNA, etc. The receptors job is to generate a quantitative signal upon recognizing the analyte...analyte. This signal depends on the system and the receptor, but readouts could involve fluorescence, pH change...

...of the Fc receptors FcγRIIB1 and FcγRIIB, inhibit) specific immune responses. These receptors are specific...so IgG isotypes bind to Fcγ receptors; IgE isotypes bind to Fcε receptors; and so on and so forth. There...of different receptors, but their downstream effects can be sorted into three main effector functions.  ... about Fc effector functions and all that antibodies can do! For instance, some Fc receptors, once bound... RA, Towers TL, Presta LG, Ravetch JV. Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor...antibody's binding affinity (Janda et al., 2016; Torres & Casadevall, 2008) but…does it really do anything... the innate immune response, by binding to Fc receptors on other immune cells. (psst! Need a quick review...

...Production at Addgene Viral Vectors 101: What is a Virus? Viral Vectors 101: Viral Vector Elements ...nucleic acid cargo of your choice. First, a viral vector is selected to perform the transduction with: either...desired DNA to be integrated can be cloned into the vector that you have selected. The nucleic acid will be...produced, and viral particles are packaged, when the vector is expressed. Once the design and construction ... it’s time to make virus! The constructed viral vector containing the cargo of interest is then typically...layers of risk management when working with viral vectors. Typically, one day after transfection into the...variety of broadly applicable and popular viral vectors. Check out our viral selections to see if we have...

...are doing is telling a (nonfiction) story. For there to be a story, you need to have conflict. There are...complete your story: an introduction, and a resolution.   Fig. 2: The structure of a story - not just...communication science. Science communicators also use the hero’s journey, stories from patients or other affected...unrelated stories that they then tuck the science into. There are almost limitless storytelling options,...pursuit of achieving knowledge. Or, as we in the storytelling business like to say, science is all about navigating...inherent tension, is interesting. It’s what drives a story forward - and this drive is so powerful that it ... communications, but they’re firmly part of the story to the scientists doing the work.  And while these...

...due to factors such as the experimental cell type and the specific genes cloned into the vector.3  2A ... the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA...for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work, and how to...get started with them. Why Use multicistronic vectors? Detecting cells that are expressing your gene,...expressing your gene of interest at high levels. Unlike vectors that express screenable or selectable markers from... the same transcript. Of course multicistronic vectors do not have to exclusively be used as a means of...

...using synthetic vectors and cultured mammalian cells. Of course, these mammalian vectors must be compatible...Instead, reporter genes are often employed to easily monitor transfection efficiencies and expression levels... mammalian vectors...

...elements for vector propogation. This post, along with a future companion post on mammalian vectors, will catch...typically found as a single copy. Unlike the ARS vectors, CEN vectors are stable without integration. Yeast Episomal...plasmid maintenence within an E. coli cell, but vectors can be widely utilized across many different cell...core replication and resistance features of yeast vectors and explain how they differ from the bacterial ...previously described. Why do scientists use yeast vectors? Yeast are eukaryotes and thus contain complex ... and yeast cells. To be useful in the lab, the vectors must contain a yeast-specific origin of replication...replication of fission yeast, Schizosaccharomyces pombe, vectors at the end.) In a previous post, we briefly discussed...

...Science Mentors series: Part 1: "What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring ...choose an advisor/principal investigator (PI) that will also serve as a good mentor. To read more about what...choosing a lab for postdoctoral work. However, selecting a lab with a PI who is a good mentor is important ...to learn about her mentoring style. How can I find out if an advisor is a good mentor? You have to ask ...on this topic), for this second entry in the “Mentoring for Scientists Series”, let’s focus on how to ...what makes a good mentor, see the previous post in this series. Check out Joanne's Reddit AMA Different...Different types of mentors for grad school vs. postdoc Choosing an advisor to complete a successful PhD is different...

...confirms the previously suggested inhibitory mechanism of locomotor sensitization and might open new ways...studies using viral vectors in behavioral neuroscience. With the ability of viral vectors to fine tune gene...all of our viral vectors blog posts Find tips for getting started with viral vectors for behavioral neuroscience...is activating, inhibiting, or modulating. Viral vectors, in contrast, are considerably more versatile and...behavioral tests, but now we’ll focus on how viral vectors have changed the scope and power of three commonly...tests To study and quantify locomotion, scientists monitor an animal’s movement in an open field box or a ...particles expressing an activating chemogentic receptor (hM3Dq) into a subset of neurons within the LC...

...reversion of this mutation by an adenine base editor would restore antibiotic resistance. To find such a protein...not require any additional inhibitor protein like UGI in CBE. Base editors for the plant genome These ...2: Plant base editor platforms and their molecular components. Adenine base editors for plant We developed... developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...smaller vectors suitable for transient assays using protoplast transformation. Cytosine base editors for ...plant base editors. Plant CBE and ABE with expanded PAM compatibility All of the base editors mentioned...require other base editor platforms. Hua et al. constructed a number of plant base editors replacing the SpCas9...

...researchers.  TALENs In 2010, transcription activator-like effector nucleases (TALENs) were engineered. TALENs... plague).  Base editors The major drawback of all the endonuclease-based gene editors (meganucleases, ...generate the prototypical base editor enzymes. The cytidine base editors (CBE) use a cytidine deaminase...the adenine base editors which can help accelerate the engineering of novel base editor proteins. She is... agricultural products. But it also made SciFi director Ridley Scott wonder what would happen if these...year after the discovery of meganucleases. The storyline involves the resurrection of dinosaurs by extracting...sequences is not ideal. For the early generation gene editors, new amino acid recognition sites had to be designed...

... these plasmids. While clearly an outlier, this story had us wondering: is there a larger trend here? ...also get many more citations This isn’t the whole story, though. It would also be great to know how much... is just a first step. There are lots of other factors that could still be controlled for. As an example...

...plasmids use Rho-independent terminators, which include naturally occurring terminators, such as T7 and rrnB,...engineered terminators come close (>95%). For most purposes, however, any common terminator will suffice...suffice. Many commercial expression vectors use double terminators to reduce unwanted translation of downstream...require binding of termination factors. Although both are termination factor dependent, Polymerases I and...Posttranscriptional Regulatory Element Enhances Expression of Transgenes  Delivered by Retroviral Vectors. Journal...of yeast terminators of varying strength. A discussion of the effects of various terminators in the context...termination and polyadenylation? The role of the terminator, a sequence-based element, is to define the end...

...outside factors that need to be considered as well. In this post, we’ll touch on antibody storage, buffer....  Storage   Once you get your exciting new antibody, the second thing you’ll want to do is store it ...what temperature to store your antibodies at, but it also matters what they’re stored in - their buffer....the way, you can put your antibody in short term storage (think days to weeks) in the 4°C. If you’ll only...primarily used in Western blots and ELISAs, cannot be stored in sodium azide at all, as it inhibits HRP. In ...excitation by a laser, which is then read by a detector panel in the machine. Fluorophores are light sensitive...sensitive, so antibodies conjugated to them need to be stored either in a dark, opaque vial or wrapped in aluminum...

...Patriarchi, T. (2023). Sensitive multicolor indicators for monitoring norepinephrine in vivo. Nature Methods...Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been ...developing and depositing two new norepinephrine indicators: nLightG and nLightR (Kagiampaki et al., 2023...White insets indicate surface expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T...nLight sensors are based on alpha-1a adrenergic receptor; nLightG contains circularly permuted green fluorescent...trialed in two-photon in vivo experiments. nLight indicators are more sensitive to and selective for norepinephrine...This is important because alpha2-AR can provide inhibitory feedback in response to norepinephrine release...

... by sharing all the incredible guest posts and stories featured in our blog in 2023. Some of these were...Information DatabaseResearcher: Amos BairochViral Vectors 101: Systemic CapsidsResearchers: Cynthia Arokiaraj...