... recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019). This... was not present (Kuhlman, SK. 2008). While theoretically this strategy should work, these vectors suffered...Callaway’s lab (Fischer, KB. 2019) set-out to systematically test the potential causes of these phenomena...expression profile at multiple steps from plasmid amplification in bacteria to transgene expression in vivo.... the level of spontaneous recombination. This indicates that the amount of leaky expression can largely...of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of scrambled cre-recombinase...expression of receptors, post-translational modifications, and other factors that AAV utilizes to gain...

...In this post in the Careers in Science Communication blog series, you’ll learn about Caitlin Runne-Janczy...to do scientific editing.” It was for Cactus Communications, a company that provides scientific editing... science communication...

...about careers in science communication. When I started my Science Communication Internship with Addgene...re interested in learning more about science communication careers, you’re in the right place. For this...this series, I’ll interview three science communicators who work in the biotech, education, or nonprofit...should be. He debated taking a position as an application support/sales representative in industry or a...job. “I am really more of a multimedia science communicator than a science writer these days. I do some ...illustrations for summarizing mechanisms and applications that are relevant to things IDT does. I also...next couple of months. This involves a lot of communication with the presenters, and editing of abstracts...

...groups have generated robust tools that have practical applications in monitoring the accidental or intentional...published their results in a paper in Nature Communications announcing a new bioinformatics tool that can...at its sequence. MIT collaboration for gene classification This inquiry started when Neil Thompson, a ...opportunity to study it and see what happens when technological change happens really fast,” Thompson says. ...essential and complementary tools in a time where biological engineering and synthetic biology is expanding...from farther away in the tree of life." Nature communications 9.1 (2018): 4425. PubMed PMID: 30356044. PubMed...the lab-of-origin of engineered DNA." Nature communications 9.1 (2018): 3135. PubMed PMID: 30087331. PubMed...

...things or communications planning.” To beef up her resume, she decided to complete a certificate in Strategic...Podcast Episode on Science Communication Check out Other Science Communication Posts Resources on Addgene.org...In this post of the Careers in Science Communication blog series, you’ll hear from Susan Keown, a staff...Cancer Research Center. Susan Keown’s science communication journey started with something unexpected: termites... time position with a Washington D.C.-based communications consulting firm, Palladian Partners. “I was...Strategic Communications and Public Relations at the University of Washington and the connections she made...team meetings and Susan is assigned to cover the Clinical Research Division at Fred Hutch, so she attends...

...short-lived biotin-phenoxyl radical in the presence of hydrogen peroxide. This radical reacts with electron-...Detecting Cell–Cell Interactions." Journal of the American Chemical Society 141.5 (2019): 1833-1837. PubMed PMID...scientists have long used antibody based affinity purification and mass spectrometry approaches. Recently however...capture these interactions in a vast array of biological contexts.  Proximity labelling relies on the ...that are tagged with biotin using avidin coated purification techniques. Due to the high affinity of biotin...challenge for classic antibody based affinity purification/mass spectrometry methods.  BioID: a promiscuous...evolution until they identified a mutant with significantly increased sensitivity that did not affect the...

...light-controlled affinity purification of unmodified proteins. Affinity purification methods typically require a protein...#1: Affinity tag purification You tag YFP and use an affinity column for purification. After binding YFP...  Light controlled protein purification with OptoNBs The first application for OptoNBs takes us in vitro...It’s time to choose your own protein purification adventure. You want to purify your favorite protein...elute YFP.  Option #2: Opto-Nanobodies (OptoNBs) purification You skip adding a tag to YFP and instead use...sound like fiction, but light-controlled protein purification is now a reality thanks to OptoNBs. Recently...response to light allows for tagless protein purification as well as reversibly regulation of endogenous...

...Extremophiles - phylogenetically diverse microbes living in extreme physical and chemical conditions like...comparison indicates that like GeoCas9 and ThermoCas9, IgnaviCas9 is a type II-C Cas9. IgnaviCas9 is most... temperature is 30°C. Find IgnaviCas9 here! What can you do with IgnaviCas9? Why would a thermostable ...deploying IgnaviCas9 without concern about its stability under harsh conditions. One example application from...Stanford University became interested in exploring biological dark matter – large tracts of the microbial tree... to be diversity hotspots containing abundant biological dark matter and so the lab organized a sampling...like YNP thermal sites - are also of practical interest since they’ve previously yielded valuable enzymes...

... to help decide whether work is suitable for publication in their journal. The early peer-review attempts...information that would prevent the study to be replicated or omission from the literature. I think that...while the wet-lab resources, like constructs or biological material, can be deposited in the specific repositories...Addgene.org Learn about depositing plasmids pre-publication Find molecular biology protocols Find eBooks ...

...an experimental system comes with fewer ethical and logistical challenges than does working with animals...Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for Applied Plant...University. Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two global stock...scientists around the globe. As a member of the Brassicaceae family, Arabidopsis is related to a variety ...preference for using animal systems to demonstrate biological concepts - and you have yourself a plant publicity...publicity crisis. The Arabidopsis Biological Resource Center (ABRC) set out to remedy that with the establishment...Arabidopsis in the classroom With funding from the American Society of Plant Biologists and support from the...

...proof-of-principle experiments that replicate current knowledge, but the sensor can also kinetically outperform current...research. This year, we put out another call for applications. We had an overwhelming response and were excited...epoxide reductase (VKOR) variants. VKOR is an enzyme critical for activating clotting factors and is the target... Lin Tian’s lab at UC Davis is to generate a genetically encoded fluorescent sensor for serotonin. According... drug adjustments. Unger ultimately hopes that medical professionals can genotype the patient and rapidly...  Michal Bernach: Developing an artificial leaf replica system to study plant-bacteria and bacteria-bacteria..., Bernach sought to develop an artificial leaf replica system. “This bottom-up approach will allow me ...

...have turned to genetically encoded voltage indicators. To expand the voltage indicator toolbox, Ed Boyden...and xCas9, and requires no additional chemicals or modifications. This versatility opens up the potential...voltage indicator Archon Article contributed by Shreya Vedantam  Listen to the voltage indicator podcast...compute using electrical impulses to coordinate emotions, thoughts, and behavior. Historically, scientists...To bring greater control to Acr activity as a biological tool, Niopek and colleagues turned to optogenetics...scientists study the electrical activity in the brain by inserting electrodes into the brain. But this process...thousands of proteins for one that can report electrical activity via imaging. They took the light-sensitive...

...fluorescence indicator of K+ for optical imaging (GINKO) Genetically encoded fluorescent indicators for imaging...comparison of a genetically encoded sensor and small molecule indicator: implications for quantification of cytosolic...Javier García-Sancho Calcium Genetically encoded calcium indicators for optical imaging (GECOs), including...and ratiometric indicators An Expanded Palette of Genetically Encoded Ca2+ Indicators. Science. 2011 Sep...Teal genetically encoded calcium sensor T-GECI Blue-shifted genetically encoded Ca(2+) indicator with ...fluorescent cAMP indicator (Flamindo2) Genetically-encoded yellow fluorescent cAMP indicator with an expanded...protein-based cAMP indicator (Pink Flamindo) Red fluorescent protein-based cAMP indicator applicable to optogenetics...

...Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods Visualization...signaling molecules and are often used in clinical applications. Diabodies - Diabodies contain two Fab fragments...this can vary significantly based on the antibody and conjugate chosen. Signal Amplification The amount ...single assay. Overview of Antibody Applications Antibody-based applications can be generally classed into ...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...processing steps, such as sonication. Denature proteins, using heat and/or chemicals, and run on an SDS-PAGE...concentrates in the sample. Controls for quantification methods Quantification methods should be run with: a positive...

...using AAV for research is that it is replication-limited and typically not known to cause disease in humans...Cap controlled by tTA-TRE amplification system Gradinaru 103006 pUCmini-iCAP-PHP.S PHP.S AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 127847 pUCmini-iCAP-PHP.V1 PHP.V1 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175004 pUCmini-iCAP-AAV.CAP-B10 B10 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175005 pUCmini-iCAP-AAV.CAP-B22 B22 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 185136 pUCmini-iCAP-AAV.MaCPNS1 MaCPNS1 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 185137 pUCmini-iCAP-AAV.MaCPNS2 MaCPNS2 AAV packaging...

...immunoprecipitation (enChIP) using CRISPR. Biochemical and Biophysical Research Communications . 439 (1), 132–136. PMID:... organisms, but modifications to various Cas enzymes have extended CRISPR applications to increasingly...takes place during the S and G2 phases. Chemically or genetically inhibiting genes involved in NHEJ or upregulating...components, like miCas9 , Cas9-DN1S , or others, can increase HDR efficiency. In most applications, a large portion...genes. Each library typically contains ∼3–6 gRNAs per gene to ensure modification of every target gene...following the amplification protocol specified by the depositing lab. After amplification, you’ll need ...heritability. Not all modifications are stable across cell divisions, and some types of modifications are more frequently...

...Double floxed soma and AIS-localized genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...-DIO-JEDI-2P-WPRE Double floxed genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...-Kv-WPRE Soma and AIS-localized genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production...179464-AAV9 pAAV-hSyn-JEDI-2P-WPRE Genetically encoded voltage indicator (GEVI) JEDI-2P in AAV production... the human brain through the development and application of innovative tools enabling large-scale real-time...or when BRAIN Initiative grants are noted in publications associated with Addgene materials. Plasmids ... 113049-AAV9 pAAV-hSyn-GRAB_DA1m express the genetically-encoded fluorescent dopamine(DA) sensor GRAB_...

...Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional...Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional...Selectable Marker PI Publication Epigenetics To make targeted epigenetic modifications, researchers have ... Gene/Insert Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used...Insert Promoter Selectable Marker PI Publication Visualize A catalytically inactive Cas9 (dCas9) fused to a...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function Genome Editing...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...

...therapeutic applications. Despite the ethical controversies surrounding non-research applications, it’s clear...just as useful (if not more so) for research applications, eclipsing past genome engineering technologies...to destroy the phages upon re-exposure. More specifically, specialized Cas proteins snip foreign DNA into...number/types of Cas proteins. Makarova et al. ’s classification defines 5 types and 16 subtypes based on shared...for CRISPR genomic editing. Genome Engineering Applications In 2012, Jinek et al. first demonstrated that... PubMed lists more than 6,300 CRISPR-related publications, many of which detail work to improve the tool...various species, as well as the development of new applications. The first CRISPR papers described two main ...

...resources for this meeting's publication, we are asking participants to share practical information in the form...permission to share (e.g., publication's permission). Connie Cepko, Harvard Medical School Ali Cetin and Shenqin...contribute to this publication. Citation information for openly shared communication: Adeno-Associated ...goal of this year’s workshop is to brainstorm practical strategies for using combinations of AAV-based...result of the meeting will be a publicly shared practical guide for all scientists in the field and every...Organizing Committee: Dr. Constance Cepko, Harvard Medical School Dr. Melina Fan, Dr. Susanna Bachle, Dr. ...Addgene Confirmed Speakers Constance Cepko Harvard Medical School Ali Cetin Allen Institute for Brain Science...