...protocol and/or import and convert it into RSpace. The reverse flow is also supported, so it’s possible...searchable and shareable, but some ELNs provide better support for reproducibility than others. Unfortunately,...connected approach in action: how the RSpace ELN supports reproducibility RSpace was designed in response... principles have been implemented by providing support for connectivity to external data sources, tools...tool like an ELN. It is nevertheless critically important to be able to conveniently reference specific ...impossible to reproduce. As the graphic depicts, RSpace supports linking to all of the most commonly used online...research captured in the ELN is incomplete. Thus, to support reproducibility an ELN should have integrations...

...detectable by two different tags: a reporter and an HA tag. The reporter comes in three varieties: HRP, Spaghetti...independent of the reporter. The only caveat is that HA staining is weaker than reporter staining. Picking...Picking the right reporter for your RANBody While all three reporters stain cultured cells and tissue sections...Monster, and Chicken IgY Domain RANbody Reporters. Reporter Detection Method Uses Pros Cons Antigens...genetically tagged and instead are conjugated to reporters (i.e. fluorescent dyes, HRP, etc. ) after purification... is a RANbody? A RANbody is a fusion between a reporter and a nanobody that’s encoded on a plasmid. A ...sections, some reporters are better suited for particular applications. Read on to learn more about each...

...tricks will apply to any biology-related customer support interaction. Before contacting customer service...information as possible about the issue. For Addgene support, make sure to mention the plasmid catalog number...have real scientists answering their technical support questions (besides Addgene, I know both Abcam and...question, and now... One of the aspects of customer support that I miss is that you’re never really sure what... response, just keep in mind that our customer support scientists wear many hats at this still relatively... customer support... technical support...

... CRISPR reporters in the near future.  SRIRACCHA (stable, but reversible, integrated reporter for assaying...histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct...Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems provide ...Recently scientists have created CRISPR fluorescent reporter assays to quantify targeted Cas9 cleavage, and...and non-homologous end joining (NHEJ). In these reporters, editing typically results in a change in the ...this blog we will focus on two fluorescent CRISPR reporter methods, SRIRACCHA and GEMCherry2. We will cover... the Piggybac transposon to stably integrate a reporter gene that contains a puromycin resistance gene...

... In this flowchart, you can see the dwindling proportion of AAV particles that make it through each part...expression. The higher the titer, the better. AAV transport and transgene expression It’s clear that we need...be required. For example, retrograde AAV gets transported up the cell’s axon. While the mechanism for this...yet fully understood, we can imagine that this transport is probably not 100% efficient. So the few AAV... particles. Limit tube transfer - the fewer opportunities your viral particles have to stick to your tubes...

...departments from technical support. Technical support careers in brief Technical support careers are open to ...technical support specialist for 4 years. Read on for insights from my time in technical support including... expertise in technical support positions. When applying for technical support positions, you should try...yourself with technical support. Even having personal experience with technical support through the customer...participated in a customer support conversation, just go ahead and contact a technical support team with a question...in a technical support role. What general characteristics do jobs in technical support have? There are... technical support roles. Examples are Technical Service Representative, Technical Support Specialist,...

...Axonal Transport It has been demonstrated that certain serotypes can undergo axoplasmic transport. These...of neurons (retrograde transport) or towards the synapse (anterograde transport), which is a possible ...retrograde and anterograde transport, but they also have similar transport characteristics (velocities...strong CNS tropism (4). If axonal transport is not desirable, transport can be reduced by injecting a smaller...and using AAVs have in common? The three most important items when looking for a house and choosing an...this mantra to AAV selection, there are three important factors related to location. Location (1): What...Traveling from the original location - Axonal Transport in the CNS What tissue? What is commonly known...

...but, in today’s post, we’ll discuss some of the important considerations you should think about when developing...ICSS can now be done with neuronal cell-type and temporal precision. In ICSS, a test chamber is used in ...interested in studying a brain region that may be important in memory, an appropriate behavioral test might...more precisely characterized with spatial (CA1), temporal (real-time inhibition), and behavioral (remote...has a bit of a steep learning curve and many opportunities for mistakes. The easiest way to learn optogenetics...differential participation in reward circuitry." Cell reports 8.6 (2014): 1857-1869. Pubmed PMID: 25242321. Pubmed...

... from rare diseases by connecting patients to important resources and promoting research for rare disease...Cancer Alliance Website.  We are proud to provide support to Drs Di Schiavi and Nabavi and will continue ...learn of future rare disease research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352...Blog Previous BeHeard Awardees Other Efforts to Support Rare Disease Research: IFOPA Donating Plasmids ...

...that company culture and enjoying your job were important parts of team buidling - they relieved some of...

...mentors from your current organization but also important to build relationships outside your organization... get started with mentoring. Read this Catalyst Report “Making Mentoring Work” to learn about what makes... it work by being an educated mentee. Other opportunities: Try virtual mentoring at Mentornet.net. This...make your mentor want to keep working with you is report back with progress. Tell them how their advice ...

...fluorescent executioner caspase reporters. Several apoptosis reporters have been developed, such as the...transfer (FRET)-based executioner caspase reporters. In these reporters, the FRET pair is linked by a caspase...FlipCherry: Red fluorescence protease reporter To add to the fluorogenic reporter toolkit, the Shu Lab also designed...novel fluorogenic reporter that has a large dynamic range and high brightness. This reporter achieves approximately...Elmore, 2007). This reporter will allow scientists to observe both the spatial and temporal dynamics of apoptosis...fluorescence emission. FRET and fluorescence based reporters however have several limitations. First, FRET ...study apoptosis in vivo scientists need a protease reporter that has a large dynamic range and high brightness...

... an odd or quirky lab story you’d like to see portrayed in LEGO form, feel free to share with us at BrickingScience...

...the community as a whole could support Addgene as we have been supporting the community. While many were...Zuckerberg Initiative for supporting Addgene by providing a generous grant. This support will enable us to continue...to coronavirus-related plasmids and to receive important coronavirus-related deposits. Our valiant lab ...We’re extremely proud of the work we’ve done to support COVID-19 research. Our collection of available ...operations for all ongoing processes are entirely self-supporting. We hire more staff as we need them to meet demand...approaching science funding organizations to request support for our operations until our normal work resumed...via Addgene. Perhaps if each of many grantors supporting research could provide Addgene with a small amount...

...SLC6A8 mutations in Creatine Transporter Deficiency Creatine Transporter Deficiency (CTD) is a rare X-... to be transported from the blood and cross the blood brain barrier using creatine transporters. However...However, mutations in the transporter SLC6A8 impede this transport and result in global creatine deficiency... the SLC6A8 gene, also known as the creatine transporter. The Association for Creatine Deficiencies (ACD...parent-let non-profit founded with the mission to support and drive research into CCDS. The brain and muscles... and induce their effector functions. It’s an important process that we need to understand so that we ...source of IL-36 cytokines in the skin making it important to study what is happening within the keratinocytes...

...issues. Improving customer support with self service We strive to make our support team available whenever...reliable Zendesk system that powers our email support, this portal houses answers to all of your most commonly...core values here is to provide unmatched customer support, as this is vital to ensuring that our services..., quickly, and this is why we have a dedicated support team that can help you through any search, ordering...an answer. Luckily, there’s a trend in customer support to address this exact problem, and it’s called ... to the repertoire of tools you can use to get support for our resources with our new Help Center! Created...the Help Center to make sure that, whatever your support needs are, we’ve got you covered! In addition to...

...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used... to study gene expression in cells. But these reporters have limited use in vivo because they are based... penetration depth as well as high spatial and temporal resolution, making it ideal for visualizing gene...expression in vivo.  But what ultrasound lacked was reporter genes that could produce ultrasound contrast. ...our lab - the Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when...5202–5209 (2017). Bourdeau, R. W. et al. Acoustic reporter genes for noninvasive imaging of microorganisms... Hurt, R. C. et al. Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors...

...real time, but each of these reporters has its drawbacks. Acoustic reporter genes (ARGs) overcome these...existing bacterial reporters Limited Depth of Signal/Imaging: Fluorescent reporters are not as well suited...fluorescent or bioluminescent reporters usually only detects the signal of the reporter and fail to pinpoint what...signal is coming from.   Acoustic reporter genes (ARGs) Acoustic Reporter Genes (ARGs) are engineered gene...imaging tools The luciferase reporter gene The green fluorescent protein reporter gene Resources on Addgene.org... researchers have developed in vivo bacterial reporters that use fluorophores and luciferases to track...these limitations by using gas vesicle reporters that are detectable by an inexpensive and widely available...

...these plasmids. This is because in 2024 certain export control exceptions were made by the US Bureau of...majority of the countries we ship to, first on a temporary and now on a more lasting basis.   Table 1: Popular...   For researchers in countries with applicable export control exceptions for rabies and VSV-G genes, ... and we strive to keep our information updated, export controls and regulations are complex and may change...

... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance... editing. eGFP reporters for single base editing by APOBEC-Cas9 Though the ACE reporter quantifies APOBEC-Cas9...DSB in their previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target...editing reporters. Image from St. Martin et al., 2019. The Harris lab used these eGFP reporters to analyze...Transient reporter for editing enrichment (TREE) The Wang and Brafman lab developed a transient reporter for...cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid delivery... functional reporter system to identify and enrich for base-editing activity The reporters mentioned above...

...Genome Institute of Singapore. Chapter 3: Biology research and resources in Singapore. Chapter 4: Life as...Chew, a researcher at the Genome Institute of Singapore who recently started his own lab. We discuss some...Principal Investigator at the Genome Institute of Singapore. His lab focuses on producing biotechnologies ... singapore...

...Problems with FRET kinase reporters Difficult to Multiplex: FRET reporters are comprised of two fluorescent...phosphatase-accessible. Kinase Translocation Reporters (KTRs) Kinase Translocation Reporters (KTRs) are genetically encoded...between the nucleus and the cytoplasm, nuclear export and import rates can influence KTR translocation. Work...least one site. Phosphorylation is particularly important for regulating signal transduction and measuring...single-cell kinase activity is FRET, but FRET reporters are challenging to design and difficult to multiplex...alternative tool with their Kinase Translocation Reporters (KTRs) whose cellular localization serves as a...multiplexed together. Challenging  to Design: FRET reporter activity depends on both the distance and the ...

...Orientation)) is a powerful tool that provides precise temporal and spatial control of gene expression in vivo...gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to easily detect opsin-expressing...- or reward-related behaviors. The high spatio-temporal precision and reversible modulation of FLEx vectors...recombinase in mouse primary afferents." Scientific reports 8.1 (2018): 7321. PubMed PMID: 29743652. PubMed...

...exist for all Addgene plasmids! In an effort to support standardized persistent identification of research...Research Resource identifiers (RRIDs) which are supported by SciCrunch. Other well known PID formats are... the journal’s specific author guidelines for reporting your materials. If you deposit a plasmid with ...plasmid URLs act as unique PIDs, we’re happy to support the RRID and compact identifier formats. Acknowledgments...

...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists...heavily on fluorescent protein genes. These optical reporters have limited deep-tissue performance, and sometimes...Mikhail Shapiro’s lab has developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression...with a resolution of less than 100 µm, the ARG reporters could be successfully visualized in vivo using...demonstrates engineered gas vesicles as the first reporter genes using ultrasound and provide a major benefit...

...remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed... it is an attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics... target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay, and are...Büttner’s lab designed a calcineurin transcriptional reporter consisting of a quickly-degraded destabilized ...panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate ...Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial...

... español! Esta traducción fue regalada a Addgene por Daniel Mejías Sánchez, ¡y no podríamos estar más ...

...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs... used to study transport characteristics of various Golgi and plasma membrane reporters as well as secretory... protein transport. Franck Perez’s lab has developed a whole collection of hook and reporter proteins ...pyralis) reporter plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system...widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been...

...the pSer-incorporated polypeptide. These three components were shown to effectively incorporate phosphoserine...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...strain for pSer-incorporation The second component necessary for phosphoserine incorporation is a modified...considerations, it is important to determine how much phosphorserine has actually been incorporated into the full-length...different levels of incorporation due to differences in stability of the p-Ser incorporated protein as well...determine the stoichiometry of phospho-incorporated to non-phospho-incorporated proteins purified from the system...pSer-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can ...

... facilitate transport: retrograde transport relies on dynein, while anterograde transport relies on kinesin...take advantage of existing transport pathways in neurons. Anterograde transport is typically used for the... transmitter synthesis. Retrograde transport is used for transport of endocytosed material or molecules... impaired, and only few reports exist about dye molecules that are transported from one neuron to another...There are several reports of both retrograde and anterograde intracellular transport of some AAV serotypes...including dendritic and axonal extensions and, most importantly, any connections the neurons make to other distant...for various forms of retrograde and anterograde transport suggest that there are several parallel mechanisms...

... whether respondents saw them as not important to very important. In addition, we also explored the barriers...”. The study also showed that It is important that an ELN supports all types of data, which enables researchers...electronic ones? It is important to know in which ways your data can be exported or printed from the ELN...project. Can you meet the ELN support team? We cannot stress enough how important this is when deploying an...only 46% could be reproduced. In a recent Nature report from researchers at Amgen, it was stated that only...through all saved data and automated writing of reports. To be able to understand the situation in depth... is likely to require on-going maintenance and support.” Besides that, academic institutions tend to think...

...conjugated with a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such...indirect and sandwich ELISA the reporter signal is directly proportional to the level of antigen in the...whereas in a competitive ELISA the reporter signal is inversely proportional to the level of antigen in the...dish and incubate it with a reporter conjugated primary antibody. The reporter reacts with a provided substrate...simplicity we will discuss the more common enzyme-based reporter methods. For enzymatic signal detection, users... sandwich and competitive. In a direct ELISA a reporter enzyme conjugated antibody binds to antigen coated...antibody first binds to the antigen and then a reporter enzyme conjugated secondary antibody binds to ...

...you think there are four important times to measure that growth rate, you report the results of all four... registered reports, the gold standard for testing hypotheses Finally, Registered Reports combine all ...Reproducibility Project: Cancer Biology, where reported methods may not include how many tests were conducted...attention on the results instead of the methods. Unreported flexibility and questionable research practices...result after conducting a replication study. An important difference between the original setting and the...genetic variation, or different handling can all be important.  If we eliminate the problems above: unclear ...research finding we can be more certain that some important, previously unknown variable actually makes a ...

... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg... termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK and... SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was first identified in...molecules. In SHERLOCK a quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable...

... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...removal of GFP from the TRICK reporter. Delayed translation is known to be important in development, and Halstead...proteins to allow translation to occur. For a TRICK reporter mRNA this displacement leads to a change in the...GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP...translation, both GFP and RFP are bound to the reporter, causing it to appear yellow. The act of translation...of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization signal...copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors cycloheximide...

...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI...Gal4 activator, bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including...to have Y2H systems that avoid transcriptional reporters altogether; In the split-luciferase system, intein-mediated...

...cuts to important research funding.  So when I saw that there was going to be a march in support of science...the rain, about 4,000 supporters of science showed up to march. We had full support of local law enforcement...proved to be the perfect opportunity to cement in my children's heads the importance of science and positively...seems to break everyday. Many researchers and supporters of science marched to advocate for scientist ...for science, and numerous other reasons. It is important to remember that we, scientists and non-scientists...°W – Vanessa Sung Hundreds of scientists and supporters in Montreal took to the streets on April 22 to...Albany, Rep. Paul Tonko also came and spoke in support of science - he's one of the few members of Congress...

...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged... The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...share your constructs from lab to lab. Promega supports the Addgene mission statement: Accelerate research...(FLuc) and Renilla (RLuc) luciferase. This is important not so much for how bright you can make a reaction...couples NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected/expressing...NanoLuc luciferase. Both GpNLuc and OgNLuc BRET reporters produced bright signals without exogenous excitation...

... to report mitochondrial turnover Mitochondrial health, transport and turnover are important features...genes, reporters, or other markers into a target genome. They did this by selecting the important elements... be imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using...usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer from Yan’s ...s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial degradation...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and organelles...channelrhodopsins. CnChR1, nicknamed Chrimson, is the first reported yellow-peaked channelrhodopsin with a spectral...

...mentor Your manager should support you, just like you are supporting your report(s). This means that you ...unique skills and strengths (you will likely have reports with different levels of experience), find out ...how to prioritize but I  just added the “most important” tasks my schedule. This was bad for 2 reasons...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...expectations, answer any questions, and (most importantly) decide on a timeline. My experience is that ...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...develop your team members by giving them new opportunities. I still need to check myself sometimes to ...