We narrowed to 1,369 results for: POR

... to display neuronal retrograde transport, AAVrg. Retrograde transport refers to the direction of movement...nuclear export. Poly(A) signal in cis The polyadenylation or poly(A) signal aids in the nuclear export of ...soma to axon) and retrograde (from axon to soma) transport. Many rAAV vectors have been engineered to express...viral element necessary for RABV trans-synaptic transport. Learn more about these and other initiatives ...the transgene. The first vector contains the 5' portion of the gene and a splice donor sequence at its ...3' end, while the second vector carries the 3' portion of the gene with a splice acceptor sequence at ...viruses from the Parvoviridae family. Anterograde transport Movement of molecules, vesicles or organelles ...

...testing multiple pegRNA designs is particularly important for prime editing. CRISPR Intereference : Target...Predicted On-Target and Off-Target Activity Another important factor to keep in mind when designing your gRNA...In addition to off-target activity, it is also important to consider factors that maximize cleavage of ...generally perform by chemical transfection or electroporation. Expression can be transient, or you can generate... to target cells through microinjection or electroporation. The target cells translate the mRNA encoding...delivery the RNP to cells using cationic lipids, electroporation, or an endosomal approach. Similar to the mRNA... breaks will be repaired by NHEJ instead of incorporating your desired edit through HDR. Base and prime...

...neural populations at distinct times. Temporal considerations Temporal precision is key in optogenetic experiments... exposed to light of the correct wavelength, the pore opens, cations flow into the cell (yellow dots),...of proteins) and sensors (genetically-encoded reporters of molecular signals). These areas have each since...specific subsets of neurons, allowing precise spatiotemporal control of these neurons by turning on and ...addition of trafficking signal from Kir2.1 and ER export signal provide improved membrane targeting 589 ...the vesicular gamma aminobutyric acid (GABA) transporter (VGAT) and ChR2 is thus expressed in all GABAergic...photoswitchable" proteins offer exquisite spatial and temporal control of protein activity. Phytochrome, cryptochrome...

...and a ssDNA-fluorescently quenched reporter is added. As this reporter is degraded by Cas12a, it will release... using a fluorescent reporter. In SHERLOCK, a quenched ssRNA fluorescent reporter is added to the reaction...insertions. When designing the donor template, incorporating mutations that prevent further Cas9 cleavage...Overview of HDR. In most applications, a large portion of the Cas9-induced DSBs will still be repaired...the desired HDR-edited allele. Therefore, it is important to confirm the presence of the desired edit experimentally...pairs to the 3’ flap, and the desired edit is incorporated into the DNA by reverse transcription. The edited...applications where orientation of the insert is important. However, type I-F CASTs favor insertion in the...

...necessary in cases where extreme specificity is important. Their epitope specificity allows binding to a...antibodies only bind a single epitope, so it is important to choose a monoclonal antibody that will recognize...using a secondary against a tag, such as His, incorporated in the sdAb. sdAbs can have weak signals due...with conjugating reactions are possible. It is important to research your reaction and buffer before conjugating... the basic steps needed for each application, important controls, and any special considerations to think...which are bacterial proteins that bind to the Fc portion of an antibody. However, they are not suitable ... assay, selection of the fluorophores used is important. It is best to inversely pair the brightness of...

...transcription of the full-length virus after it is incorporated into a host genome. This is often found in second-generation...cells, they kill off any cells that have not incorporated the lentiviral genome and the cells that survive...the only type of selectable marker. Fluorescent reporters, such as GFP, are another common selection marker...interest and contains the sequences that will be incorporated into the host cell genome but cannot produce...after one round of reverse transcription. After incorporation into a host cell, this deletion prevents further... expression of transgenes via increased nuclear export. RRE in cis Rev Response Element; the sequence ...partially spliced transcripts to facilitate nuclear export. Provided by a separate plasmid from gag/pol in...

...control expression of a chemogenetic reporter ensures that the reporter remains silent until a cell or tissue-specific...ligands can have high potency, which is potentially important for dosing in animal studies, made them attractive...agonists for PSAM4. A given PSAM is coupled with an ion pore domain (IPD) to form ligand-gated ion channels (...signaling. PSAM-based LGICs can be classified by the ion pore domain they are coupled to: PSAM-Gly PSAM-Gly LGICs... of PSAMs and their activity in neurons PSAM Ion Pore Domain Ligand(s) Effect Outcome (in neurons) Reference...

...Common Bacterial Promoters Researchers often incorporate the following promoters into plasmids to drive...transcription of a given gene. There are three main portions that make up a eukaryotic promoter: the core promoter...make up the transcription complex. The most 3' portion (closest to the gene's start codon) of the core...transcription factors bind. Distal Promoter The final portion of the promoter region is called the distal promoter...Common Eukaryotic Promoters Researchers often incorporate the following promoters into plasmids to drive...

...cells, they kill off any cells that have not incorporated the retroviral genome and the cells that survive...the only type of selectable marker. Fluorescent reporters, such as GFP, are another common selection marker...interest and contains the sequences that will be incorporated into the host cell genome but cannot produce...after one round of reverse transcription. After incorporation into a host cell, this deletion prevents further... expression of transgenes via increased nuclear export. Psi (Ѱ) in cis RNA packaging signal; recognized...

...can help you begin to understand what genes are important to a certain phenotype, and allow you to design...the same basic process: Amplify the library (electroporation and maxiprep). If delivering as virus, make...library to cells. The screens diverge at a few important points, including when to apply selection, when...die or do not pass selection (in the case of a reporter) Sort “winning” cells Sequence those cells that...

... and ligated into your desired plasmid. It is important to note that restriction enzyme target sites can...throughout endogenous DNA sequences. Thus, it is important to confirm that there are no additional sites ...replication process will repair the nicks. It is important to note that LIC has difficulty assembling DNA...

...first-generation rAdV vectors than later generations, which incorporate additional genomic deletions. Figure 2: First-generation...Adenovirus Vector as a Zika virus vaccine . Scientific Reports, 10 (1), 3613. https://doi.org/10.1038/s41598-020...adenoviral genes provided in trans . Seroprevalence Proportion of individuals within a population that present...

...Plasmids with control elements and fluorescent reporters to be used in combination with the MoClo-YTK toolkit...

... 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...

...everywhere can access plasmids despite the evolving export/import rules. Our shipping and lab teams continue to...entire supportive Board, we have a robust committee process in place and continue to incorporate additional...all of the success we have achieved and the opportunities we have for the future. At Addgene we will never... The success of these projects allows us to support more new projects. Diversification is happening... quality and success. Customer and technical support Our customer and technical service brands continue...sharing, but we don’t have to worry about getting support from depositors. We regularly hear how much they...blessed with a large community of scientists who support us by depositing, requesting and spreading the ...

...plasmids, are export-controlled materials in the USA, where Addgene is located.  The export license, if ...requestors. We also offer export license applications for VSV and SARS-COV-1 plasmids. Export control license ...Addgene is excited to announce we now offer an export license application for rabies materials! Some ...license process    An export control license, once approved, allows Addgene to ship plasmids or viral vectors...the applicable research project when purchasing export-controlled materials. Addgene will then apply for...process.   Due to recent regulation changes, an export control license for rabies materials may be required...required for others. You will be notified if an export license is not required for your country after ...

...STEAMid. Internships open doors to a web of opportunities that the classroom cannot offer. Furthermore... not deter any student from applying to the opportunities of their dream.  To help students focus on the...below:  1. Eligibility check Scrutinize every opportunity prior to starting an application to assess your...unless you are applying for a Federal Government opportunity, which tends to be more detailed. Beyond the ...draft a personal statement, which is the perfect opportunity to paint a comprehensive picture of your academic...statement will pay dividends. This is the only opportunity where you get to impress the judges on your ability...your recommenders deliver a superb and detailed portrayal of yourself. Be mindful that January and February...

...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events..., D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions...FPs) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically. Fortunately...should be supplied (Baloban et al., 2017).  It is important to consider effective brightness of NIR FPs in... a cell and depends on the efficiency of BV incorporation by an apoprotein and protein expression level...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...Oliinyk et al., 2019). They were engineered to incorporate BV. Their advantages are their small size (17...

...Figure 1). Addition of substrate generates a proportionate luminescent signal, strong enough to detect ...signal to gradually diminish over time; and, importantly, repeated excitation generating reactive oxygen...targets in translational research Researchers can incorporate HiBiT with a wide range of workflows, from classic...paper for more detail on whether the authors incorporated HiBiT by CRISPR tagging at the endogenous locus...the authors used a p53 response-element NanoLuc reporter to show that the mutants abolished transcriptional...editors Tracking factors that utilize exosomal transport is quite challenging. Work by Shibata et al. (...internalized. Functional assays further showed that imported ADAR1 increased oncogenic editing of its most ...

...sequence are important considerations for designing your gRNAs. For indels, it's not so important what location...nom. Me love gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment ...tool among them. While this seems obvious, it is important to remember that the same is true when designing...location in the gene you target, but it is important that your gRNA sequence is designed to be highly active...CRISPRi, these considerations are of roughly equal importance (target should be near the TSS but you can worry...from). Finally, for HDR, location is much more important because you have to target within ~30 nt of your...gRNA with an optimized sequence is of primary importance (more on this below). Created with BioRender.com...

...Antibody Data Hub. Data reports  Each antibody submission is used to create a data report, which contains experimental...that information in the report as well.    Figure 3: An antibody data report, showing the results of ...methods section for the data report shown in Figure 3.   At the end of the report, you’ll find a results section... the data report shown in Figure 3. The antibody is rated "Pass". Curating the data reports Just like... page of the Addgene Data Hub.   Antibody data reports can be searched and sorted by gene/target, name...information is then formatted and published as a report with a DOI for citation. If you’re considering ...

...Career Development; a Stakeholder Workshop Report.” The report enabled the development of priorities for...programs are still doing an inadequate job of supporting science trainee transition into diverse, successful...amplified by systemic biases which, in the absence of support and structure, prevent science from attracting ...Development Hub Workshop Attendees and their 2021 report. Image from pd|hub.   To begin to tackle these... of Massachusetts Medical School. It has been supported by a Steering Committee and working groups with...the findings were captured in this just released report “Enhancing Dissemination of Evidence-Based Models...Institute of General Medical Sciences (NIGMS) will support the development and implementation of collections...

...variety of pore sizes. If your protein of interest is small, you may want to consider a smaller pore size (...primary antibody required. Reporters There are two main classes of reporters: chemiluminescence enzymes...protocol and protocol video!  Antibodies The most important part of your western is your antibodies, particularly...you need a specific conjugate, and changing your reporter molecule requires either re-conjugating or re-...-purchasing your primary antibody with the new reporter. Finally, each new conjugate will have to be tested... weight proteins Long running time   It’s important to note that Bis and Tris gels run proteins in...high protein retention. It comes in a variety of pore sizes and protein capacity, allowing you to optimize...

...fluorescent reporter, directly couple silencing of your gene target to silencing of the reporter. This is...application of long or short dsRNA. However, it is important to note that naked and unmodified dsRNA elicits... Together, this excellent work highlights the importance of using orthogonal methods to confidently conclude...fusing your gene target sequence to a fluorescent reporter sequence (discussed below). Examples of these ...genome, whereas shRNAs are exogenously derived. Importantly, instead of the assortment of siRNAs generated...contains a target sequence against the fluorescent reporter in an inverted repeat orientation. Therefore, ...sequence against your gene(s) of interest and the reporter. This approach greatly enhances experimental throughput...

...governments. Most communities have opportunities to join commissions or boards, support ballot initiatives, deliver...are ways you can hone important skills for the profession. One of the most important skill sets for a science...success in a science policy career with many opportunities to pivot and grow along the way. That’s what...fellowships, volunteer positions, and part-time opportunities. But even through all my roles and my current... of the who, what, where, when, why and most importantly, how, of a science policy career.  Who does science...science policy? I would argue that this is the most important question that you can explore. The answer will...advice for you and your next steps. This advice is important, but must be taken through your own personal lens...

... world. To support this pace of research, new, centralized databases and centers to support the sharing...like here at Addgene!), and new databases are supporting their access to useful resources during this ...information about reagents, funding and volunteer opportunities, protocols, publications, and more to accelerate...the COVID-19 Resources Canada site. To date, the portal has recruited 3,218 Canadian volunteers, and features...COVID-19 related research projects, funding opportunities, and community response initiatives. Accelerate...research Universities have also been stepping up to support COVID-19 research. The new Center for COVID-19 ...Preparedness (CCRP2) at Rutgers University not only supports collaborations at Rutgers, but also lists data...