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TypeBlog Post...Storytelling is a crucial part of interpersonal communication, which makes honing your ability to tell stories...Addgene Blog Check out our blog posts on science communication Find interviews scientists in SciComm careers... science communication...
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Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools
TypeBlog Post...makes them more adaptable to a broad range of applications. Recently, the Spiess lab developed a set of... with specific genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered...only 1 min. Using these lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed... -
Multifunctional Neural Fibers: Modulation from Gut to Brain
TypeBlog Post...channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused to mCherry to the brain tissue. ... several months after implantation of fibers, indicating the system was indeed stable for continuous monitoring...interfaces with the entire body, so the obvious next applications are to expand the tool to different brain-organ... -
Antibodies 101: Epitope Availability
TypeBlog Post...Conformation changes usually fall into one of two categories: ones induced by the researcher and ones that...naturally, by binding to other proteins, changing locations, or even just as part of performing a specific...it a transmembrane protein, or is it know to translocate or fold in certain conditions? It can help to... -
Plasmids 101: A Brief History of Plasmids and an Improved eBook!
TypeBlog Post...clarify the classification of these cytoplasmic inheritance factors. He proposed the catch-all term “plasmid... as bacterial conjugation and recombination, replication and topology, and cloning and gene expression... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...non-trivial, however, as FPs are tremendously complicated entities with a large range of characteristics...decision remains spread across literature in publications that introduce these tools. Years ago, Kurt...incorporate FPbase data in their own projects and applications, a REST API allows programmatic retrieval of... -
Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System
TypeBlog Post...research. This year, we put out another call for applications. We had an overwhelming response and were excited...sensor with proof-of-principle experiments that replicate current knowledge, but the sensor can also kinetically...fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be diluted out and ... -
Predicting Adverse Reactions to Monoclonal Antibody Drugs
TypeBlog Post...silico approach The in silico approach cannot replicate the complexity of antigen processing events that...This system would somewhat act like a 2-step verification. All approaches have their own limitations. ... together can inform decisions about in vivo application and clinical testing. This would pave the way... -
K. phaffii: Rising to the Occasion
TypeBlog Post...2017). Beyond human relevance, yeast have many applications in food science and biotech due to its fermentation... ancient yeast strains and metazoan cells. Applications for K. phaffii There’s a number of different...is exceptionally good for post-translational modification of recombinant protein products, specifically... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog Post...Glutamate plays a variety of roles in synaptic communication and can trigger other forms of neuronal signaling...SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate Indicators for Synaptic Imaging However, the first two generations... 30377363 Aggarwal, Abhi, et al. . “Glutamate indicators with improved activation kinetics and localiation... -
Fluorescent Proteins 101: Luciferases
TypeBlog Post...The North American firefly, Photinus pyralis, catalyzes a chemical reaction in its abdomen to produce ... Figure 2: Simplified chemical reaction catalyzed by luciferase, releasing light. Created with BioRender.com...NanoBiT, are expressed in two subunits and only catalyze a luminescent reaction when they come together... -
Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors
TypeBlog Post... specific DNA base changes and consist of a catalytically impaired Cas protein (dCas or Cas nickase) fused... SpCas9. In order to expand the gene-editing applications and targeting scope of ABEs David Liu’s lab ...through many rounds of mutations, selection, and replications per day. The PACE and PANCE systems use M13... -
Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection
TypeBlog Post...historical standard. Specifically, in our recent publications, we used the cancer plasmid collections listed...pathway reference collection is described in this publication We also wrote an extensive review on the exciting...data generated from these alleles in the above publications), it will be possible for many others to perform... -
Screening for Successful Genome Editing with Digital PCR
TypeBlog Post...a gene of interest is one of the most common applications of precision genome editing, the remainder of...single molecule resolution of digital PCR allows classification of individual target molecules based on their...single wells of a 96-well plate without prior quantification. Comparatively, mismatch assays require between... -
A Novice's Perspective on Neuroscience - SfN2017
TypeBlog Post...the things that popped out to me as a novice and indicate why other neuroscience newbies might want to get... reinforce our goals of making more AAVs and educational content to help researchers use these great tools...start making the AAV, or help us produce new educational content on the blog. Overall, there are many ... -
Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection
TypeBlog Post...Scientists can also use electroporation, the application of an electrical charge to cells, to increase... properties. The phagemids are packaged into replication-incompetent phage particles with assistance from... found on an episome, a piece of DNA that can replicate on its own or be integrated within a bacterial... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog Post...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...Hohlbein, et. al., 2022). A more open way of communicating in science brings stronger networking across... -
Building and Validating Recombinant Antibodies: A Case Study
TypeBlog Post...cDNA, which was used as a template for PCR amplification with a degenerate primer set to amplify IgG ... While screening by IF-ICC worked well for identification of the correct clones, it was determined that...through functional assays. Following functional verification, the R-mAb expression plasmids were DNA sequenced... -
Antibodies 101: Monoclonal Antibodies
TypeBlog Post...cells and antibody-producing B cells that cannot replicate, meaning that only hybridomas will survive this... Antibodies: Distinguishing Characteristics, Applications, and Information Resources. ILAR Journal 46:... the Addgene blog Learn more about research applications for antibodies in Intro to Antibodies Read about... -
Kiran Musunuru on the Newest TALEN Genome-Editing System
TypeBlog Post...Musunuru: The kit includes 832 plasmids containing prefabricated pieces that allow you to build a DNA-binding...diseases. We are knocking out genes that have been implicated in disease or, in cases where particular variants...less than a month's time. Addgene: What are the implications of this kit in terms of understanding human ... -
Addgene: The First Twenty Years
TypeBlog Post...as well as developing and sharing supportive educational resources. By 2017, we had distributed 1,000 ...We also created a growing library of antibody educational resources, including our new Antibodies 101 eBook...community by depositing, ordering, or using our educational resources. And we look forward to twenty more... -
CRISPR 101: Cas9 vs. The Other Cas(s)
TypeBlog Post...a defined site to form a clean DNA break, Cas3 locates its target sequence and initiates single stranded... out large regions of DNA and has anti-viral application potential as a result. Fast facts – PAM requirement... has been proposed to be useful for the same applications (RNA knockdown, RNA editing, etc.). The main... -
Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening
TypeBlog Post...sequences via oligo synthesis using the format indicated below and pool-clone them into the pAWp28 storage...realization of a variety of perturbations and applications in functional genomics, cell reprogramming, ...CRISPR/Optogentics Tools Resources on Addgene.org Catch up on Your CRISPR Background with Our CRISPR Guide... -
CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9
TypeBlog Post...it is important to carefully select the target locations of knock-in studies to avoid off-target effects...it comes to targeting precision and practical application. If the CRISPR/Cas9 system is not effective for...lncRNAs with high specificity, particularly in applications requiring smaller delivery systems (Zibitt et... -
Making CRISPR Plasmids Using Fragmid
TypeBlog Post...ampicillin resistance cassette and an origin of replication, which are not schematized. Sizes of fragments...five months. When they tested those clones (in duplicate), 93% had the correct banding pattern in the restriction...correct. Because they created the assemblies in duplicate, they were able to confirm that at least one plasmid... -
Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog Post...passionate about HIV cure research and science communication. References 1. Gaspar, H. B., S. Howe, and...affinity-purified receptor-targeted viral vectors." Nature communications 6 (2015): ncomms7246. PubMed PMID: 25665714...our AAV Production Protocol Check out our AAV Purification Protocol ... -
Popular Retroviral Vectors and Their Uses in Scientific Research
TypeBlog Post...-humanized (Plasmid #44246) Expression of a catalytically inactive (dead), human codon-optimized Cas9 ...be used to construct complex lentiviruses for modification of mammalian cells with a single viral infection...Gammaretroviral Vectors: Biology, Technology and Application. Viruses. 2011. PubMed PMID: 21994751. PubMed... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog Post...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...BioRender.com In their recent ACS Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive...regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes bringing the C-terminal... -
Plasmids 101: FLEx Vectors
TypeBlog Post...sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability of... the function of various point mutations and truncations in the absence of the endogenous gene. Spatio-temporal..., allowing for clearer characterization of the location, morphology, and circuit mapping of a targeted... -
How-to: 5 Steps to a Great Panel Discussion
TypeBlog Post... about making sure the audience is interested, educated and talking as a result of the content. Here are...contribute. The moderator should identify the funny, provocative or interesting stories that will make for a fun...sure all your panelists have the correct date, location and parking information and remind them about ...