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...post are engineered to bind RNA sequences derived from Zika virus and, upon binding, produce reporter proteins...posts written by Addgene depositors. Below you’ll find brief summaries of several excellent guest posts... post, Luke Lavis from Janelia Research Campus reminds us that, although fluorescent proteins are incredibly...used in a wide variety of experiments. Read more to find chemical fluorophores for your next experiment. ...virus biology while leaving virulent components behind. Minigenomes are stripped down viral genomes containing...post for potential applications of R-bodies and to find a video of them in action. AAV: A Versatile Viral...Medical School describes how AAVs function and indicates important things to consider if you’re getting...

...Addgene.org Find Photoactivatable FPs in Empty Backbones in the Michael Davidson Collection Find Photoactivatable...from a dark state to a bright fluorescent state (“kindling”), and from a bright state to a dark non-fluorescent... In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't become inactivated...photoactivation. A high contrast ratio usually indicates that a PA-FP has a low level of spontaneous fluorescence...photobleaching and phototoxicity that standard FPs might induce though their continuous excitation. By generating...photoswitching between two fluorescence wavelengths. Indeed, within the world of super resolution microscopy...localization information in cells. A point to keep in mind- although PA-FPs are vastly improved over the conventional...

...proteins that bind the RNA motif will be biotinylated. Read more about RapID in this blog! Find plasmids for...biotin. Biotin, is a natural coenzyme, that strongly binds the glycoprotein, avidin or a similar protein, streptavidin...biotin molecules to dissociate from the ligase and bind to exposed lysines on neighboring proteins. Thus...organoid or in vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development ...biotin ligase from A. aeolicus, which lacks the DNA-binding domain (233 amino acids) found in the E. coli version...fusion proteins to specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate...Lam et al. 2014). They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid ...

...works GPCR signaling is initiated when a ligand binds to the extracellular surface of the GPCR. This results...from the Gβγ dimer. The Gα and Gβγ subunits then induce or inhibit intracellular signaling cascades as .... Ligand dissociation from the GPCR allows for binding of a new inactive heterotrimeric G protein complex...identify some of these unknown ligands and their GPCR binding partner, Bryan Roth’s lab created an open source...activation for more than 300 GPCRs. In this assay, binding of an agonist recruits a beta-arrestin-TEV protease... discover new ligands for unknown GPCRs. You can find the entire PRESTO-TANGO kit at Addgene! TRUPATH:...were first mutated via site-directed mutagenesis to bind non natural ligands. These non natural ligands however...

... PhDs Making the Leap to Industry." He successfully transitioned to industry after his PhD and a couple... PhDs face when transitioning from academia to industry is whether to stay hands-on in the lab or pivot...exploring alternative career trajectories within the industry, but the transition wasn’t easy - I had to reinvent...assistant. However, I recognized the need to shift my mindset to empower my team and allow myself to focus on... rather than the day-to-day details. Ignoring individual differencesManaging a team isn’t “one size fits...a diverse team where some members thrived on independence, while others required more guidance and structure...learned that my approach needed to adapt based on individual needs. For instance, I had one colleague who ...

...question after credible attempts to replicate major findings could not be replicated by other researchers.There...replicate.  Why is replicating previous research findings so difficult to do?  There are four reasons that... to present only the most novel or significant findings, and combined with powerful cognitive biases such... such as motivated reasoning and hindsight bias, we may fool ourselves and our readers into believing ...research practices (QRPs) can quickly lead us to find statistically significant results that are not robust... not come out the same as an original research finding we can be more certain that some important, previously...makes a big difference for the robustness of the findings.   How do we make research more reproducible?...

...100% indel frequency at both targets (35% and 76% biallelic), compared with only 44% and 66% indel frequency...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...cleavage, they examined gRNA expression levels and indel generation for two PTG constructs in rice protoplasts... allowing them to examine the frequency of both indels at each locus, as well as large deletions. They...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct....1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication at Addgene ...

...interest flowed through. The protein of interest binds to the metal ions, becoming immobilized itself. ...containing resin. The protein of interest (purple) binds to the Ni2+ and is immobilized along with a polyhistidine...purification, however, is the propensity for nonspecific binding of untagged proteins. Despite being a relatively...histidines are present in most production systems and can bind and co-elute with the protein of interest (Bornhorst...be high enough to disrupt contaminant-metal ion binding without eluting the target protein and needs to...substrates along with stringent wash conditions.   Find polyhistidine tag plasmids here! Where should I ...Unlike other affinity purification methods where binding relies on a specific protein conformation, the ...

...those of antibody-independent assays such as mRNA data from transcriptomics; independent antibody strategies... internet search for the required antibodies and find….hundreds of options. With limited time and budget...immunoprecipitation, the epitope may not be accessible for binding and the antibody will not work. Whenever possible...more detailed data on the assay methods. You might find that while the website highlights a variation of...fluorophore, or a conjugated secondary antibody that will bind to your primary antibody. If you are trying to visualize...applications use either direct (primary-conjugated) or indirect (secondary) methods. In those cases, remember ...the antibody cross-reacts with. Ideally you will find an antibody that has been validated in the species...

...small molecules that bind these target proteins to other small molecules that bind an E3 ligase. These ...molecules by conjugating a CRBN-binding ligand (thalidomide) to an FKBP12F36V binding ligand (ortho-AP1867) via...E3 ligases such as cereblon (CRBN) or von Hippel-Lindau (VHL) to bring the endogenous degradation machinery...selectivity, overcoming resistance to inhibitors, and inducing prolonged biological effects. While targeted ...oncoproteins that lack selective small molecule binders. To overcome this limitation, we created a generalizable... FKBP12F36V-tagged proteins. The dTAG molecule induces ternary complex formation between the FKBP12F36V-fusion...and the ubiquitin proteasome machinery through binding with cereblon, leading to rapid target degradation...

... articles. Here's what you'll find in this post: Glutamate indicators with increased sensitivity and...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation... for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean,..., Iyer, R., … Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation...and AAV plasmid sets launching later this year!  Find Addgene's own Lentiviral backbone plasmids here!... 5% and 11%. It has higher efficiency and lower indel formation than methods relying on homology-directed...scarless, unlike previous integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger, L...

...options critically. How will it feel when you find it? If you find the right lab, some days it won’t even feel...scientists seem to toss data-driven reasoning out the window when making decisions. We think scientists make...success. Gather data on the lab choices How do you find out what the lab is really like? Gather data of ...boss You sign on to a lab for the mentoring only to find out that the lab head is an absentee advisor. They...character traits and skills that you hope to grow.  Find mentors who take their responsibility to train you...meaningless tasks, removal of responsibility, repeated reminders of blunders, setting target up to fail, shifting...(topic and/or technical aspects) applicable to industry if this path interests you  (e.g. human disease...

...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...deposit please sign up here.  Here's what you'll find in this post: Tools for labeling excitatory inhibitory...respectively. Once expressed in the neuron, the FingRs bind to the endogenous synaptic proteins allowing them...inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration ... show a high proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in ...in high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...a few highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene...

...at Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET... particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation...directly interact with the bait from those that indirectly interact or from proteins that are just in the...labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein and saturation...Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting proteins... PMID: 28585547. PubMed Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann...fragmented at a different site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim...

...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find new CRISPR tools for your research! This time: ...control. Read the paper in Nature Communications Find the E. coli genome-wide CRISPR inhibition library...fold. Read the paper in Communications Biology Find the Cas9-PCV and PCV-Cas9 plasmids Selective curbing... editing efficiency. Read the paper in Nature Find the SECURE base editors All-in-one AAV delivery... in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR interference...essential C. albicans gene. Read the paper in mSphere Find the dCas9 and dCas9-Mxi1 plasmids If you have a...

...most prepared of us can find ourselves talking to our posters, so if you find yourself in this situation...session, or it may be held at an unfamiliar venue. Finding your poster board well ahead of time ensures you...at the actual venue. Introduce yourself: I always find it odd when I walk up to someone’s poster, and then... a lap around the posters and read the titles to find the ones you’d like to revisit. Strategically traverse...give the “elevator pitch,” of his/her project – kindly let the presenter know you are very interested ...either seek out the presenter after the session or find him/her on social media or through their affiliations...who presented on a topic related to your research? Find a way to get in contact with the presenter after...

...specific epitope that your antibody binds. For example, Myc1-9E10 binds EQKLISEEDL (pronounced “equi-kli-...Resources on Addgene.org  Find recombinant anti-c-Myc [9E10] at Addgene Find plasmids with the myc-tag...same antibody developed almost 40 years ago? Well…kinda. In some cases, antibody providers are distributing...to protein engineering. As a result, you can now find Myc1-9E10 in a variety of isotypes like rat IgG1...this trusty tool doesn’t end there: you can also find anti-Myc [9E10] Fab fragments and single-chain variable...accessibility and the variety of formats you can find it in likely mean it will remain an important tool...tool for the foreseeable future. These newer findings simply add to our understanding of how this antibody...

... weak gRNAs (CFD < 20) bind almost not at all, and strong gRNAs (CFD > 80) bind almost as much as perfect... when designing gRNAs Find other CRISPR software Resources on Addgene.org Find CRISPR plasmids and resources...rest of this article, we'll explain the thinking behind the tool, its key features, how it works, and how...we began using it internally. Guiding principles behind CrispyCrunch While CrispyCrunch may evolve over... not available publicly. Furthermore, we did not find any tools which were optimized for HDR, except for...typically advised to mutate the PAM to which the guide binds. However, if the PAM resides in a coding region,...single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR may inadvertently...

...survey. When asked if they could detect indels (an indication of gene disruption ability) with NgAgo, ...surveyed here could not achieve indels with the NgAgo system indicates that it is very tricky to use and...many researchers were having difficulty achieving indel formation (gene disruption) with this new technology..., one respondent out of 88 confirmed successful indel formation at a target site. Also, 41 respondents...dead in the water. On the bright side, successful indel generation and epitope knockin was achieved by a... a few respondents suggesting that NgAgo can indeed be optimized for genome editing in mammalian cells...transfections with the ssDNA guide, although many may find this inconvenient. Another possible reason for the...

...interference, can reversibly exert their effect independent of gene copy number. RNA interference (RNAi) ... 1). As a result, gene silencing can be easily induced by exogenous application of long or short dsRNA...applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi...experiments that require exogenous protein, successful induction of RNAi only requires the presence of dsRNA. In...phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long hairpin RNA.... Each construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene...more specific than long hpRNAs, they must be individually engineered and empirically evaluated for their...