We narrowed to 614 results for: cat.1

...participants could share their notes together. Figure 1: Can you draw Spongebob from these instructions without...session to another,” Bachle says. “Now, the physical location didn’t matter. I didn’t have to consider that ...

...vector-exosomes), exosome AAVs, or exo-AAVs (Figure 1). The researchers discovered that these particles ...the classical purification protocol involving iodixanol gradients. The current purification protocols for...particles accumulate inside the producer cells and purification protocols often describe a process that begins...lysis to release AAV particles with subsequent purification steps. It’s also possible for AAV vectors to... This is particularly useful for therapeutic applications in vivo where endogenous anti-AAV antibodies...Adrienn, et al. "Virus vector-mediated genetic modification of brain tumor stromal cells after intravenous...

...pitfalls? Here's what you need to know.   Figure 1: Management can seem a mystery at first! Image used...cohesive team environment. Neglecting communication skillsClear communication is the backbone of effective management...management. Miscommunication often arises when managers fail to set expectations or provide timely feedback...firsthand when it took me some time to adapt to the communication style of a new, junior team member who had values...and leveraging tools like the Myers-Briggs Type Indicator or DISC assessments can help you navigate team...

...the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two MS2, PP7, or boxB sequences...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...has been widely adapted for a wide variety of applications: base editing, CRISPR activation and inhibition... (B) CRISPR-Sirius used to visualize relative locations of the subtelomeric region of the pericentromeric...al. “CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging.” Nature Methods 15 (2018)...

... genes with a single viral vector.   Fig. 1: Schematic of multiplex gene editing with DAP array...packaged into viral vectors for potential in vivo applications or multiplex CRISPR screening.   Using tRNAs...multiplex genome engineering of crops or allow for sophisticated CRISPR screenings that would perturb the unprecedented...and efficient in plasmid and viral vectors. In applications when repetitive tRNA sequences must be avoided...construct DAP array for multiplex gene editing applications, we provide detailed manual and online design...

...as an endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head ...antibodies may just not perform as well in one application as another, so it is always best to have options...for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page for more...fluorescent proteins (FPs) necessary for many of these applications have lagged. Fixation, chemical staining, and...well-suited for the demands of advanced microscopy applications such as super resolution, electron, and expansion...

...databases, EENdb (published 1 Jan 2013, no longer available), is a simple catalogue of reported sgRNA designs...follow him on Twitter @CMacPhD.          References  1. Ghorbal, Mehdi, et al. "Genome editing in the human...into this category describe how the user is expected to interact with the software. This category is important... CRISPR software submissions. With few prior publications to go on, a submitted manuscript could only ... solutions that facilitate most experimental applications in CRISPR engineering. When compared to that...easy to fault the lack of features in a single application. The question is then, should we use all tools...novo sgRNA design, I again separate it into two categories. On the one hand we have software tools that ...

...settings with experimental samples. Figure 1: Example flow plots for a single color control. A)...ll need a few controls specific to flow as an application. These controls will allow you to distinguish...commercial antibody pages. Flow cytometry can be a complicated and daunting process. Controls like single color...

...and also by a lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate...insert. All colonies on this plate should be blue, indicating that your IPTG and x-gal are working as they ...of false positives: Blue-white screening only indicates the presence of AN insert, not necessarily YOUR...

...Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One of my favorites is making the...great way to get started with mentoring. Read this Catalyst Report “Making Mentoring Work” to learn about ...tapping into it and helping make it work by being an educated mentee. Other opportunities: Try virtual mentoring...

...usually 1-3 of the first 3 bps are modified) for optimal stability and efficacy.    Base modifications What...switches. Stabilizing gRNA modifications  Sugar phosphate backbone modifications The recognition sequence...phosphorothioates with 2’-O-methyl modifications, proved more stability than just one modification alone (Hendel, et al...ordered as synthesis modifications when purchasing your gRNA. Pro tip: modifications are typically incorporated...stabilizing modifications listed above were developed before CRISPR-Cas9, for use in applications like siRNA...worrying about the status of your RNA. gRNAs modifications are even going beyond stability, adding color...cellular RNA molecules with post translational modifications. One of the most common of these is 2’-O-methylation...

...medRxiv.org) in the past few years.   Figure 1: Stand aside, CERN — it's a new age of accelerating...their CV or grant applications. A number of organizations (such as ASAPbio) advocate for preprints, and...the most important part of your research is communicating the results of the work. Yes, those control ...any impact! There are lots of ways scientists communicate with each other: from formal mechanisms like ...poster or talk, or submitting patent or grant applications; to informal settings like lab meetings, social...of the most significant shifts in scientific communication is the rising use of preprints, or scientific...publishing in a journal.  Accelerating scientific communication Preprints have been common in fields like math...

...antibody that doesn't work for your application.      Fig. 1: Antibody Selection Flow Chart  ...your specific application. Consider also that there may be variations within an application. Take immunohistochemistry...website highlights a variation of your application, a publication successfully used it in a similar way...choosing an appropriate antibody first look at the application-specific data provided on the manufacturer’s ... a linear antigen tend to work very well for applications that denature a protein to its primary (linear...the most similar to yours. Take a look at the publications cited on the manufacturer’s website, which may...plan to. To secondary or not to secondary?  The application will also determine whether you should use an...

...techniques?  Let us know in the comments! References 1. Bacteriophage P1 site-specific recombination. I. ...recombinase catalyzes the site specific recombination event between two loxP sites, which can be located either...maintained).  Translocation: If the sites are on separate DNA molecules, a translocation event is generated...important plasmid elements – promoters, origins of replication, protein tags, and antibiotic resistance markers...creating (excuse the pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells,...proteins or co-factors, thus allowing for broad applications in a variety of experiments. LoxP (locus of ...and the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT. The loxP sequence does not occur...

... contact her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization...This application of AHA labeling is known as fluorescence noncanonical amino acid tagging (FUNCAT, Dieterich...experiments. Visualizing newly synthesized proteins with FUNCAT Professor Schuman’s technique applies metabolic...the whole de novo synthesized proteome. However, FUNCAT alone cannot be used to determine the turnover ...specific protein. Professor Schuman’s lab showed that FUNCAT’s usefulness can be extended to determine the turnover...ligation assay (PLA). The clever combination of FUNCAT and PLA PLA was originally published by the Söderberg...DNA strands take part in rolling circle DNA amplification. The amplified DNA segment is labeled with fluorescent...

...efficiently with adenovirus and AAV.   Table 1: Types of viruses and their features Choosing ...injections, or the location of a specific cell type doesn’t always lend itself to direct application of virus. ...addition of a helper virus or plasmid allows it to replicate episomally, essentially remaining an extrachromosomal...extrachromosomal part of the host genome that can replicate and divide along with the endogenous DNA. AAV also...gamma-retro viruses. This is large enough for many applications, but in some cases multiple AAVs (co-infection...and AAV. If you’re wondering more about what applications these viruses are good for, we have plenty of...Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New...

...advice below:  1. Eligibility check Scrutinize every opportunity prior to starting an application to assess...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources... detailed. Beyond the resume, a cover letter communicates your interest in a specific program, and institution...February represent peak season for internship applications. Thus these noble individuals are inundated ... with a stellar letter of support.        5. Application submission  Set a calendar reminder for every...about outreach among low-income communities to educate parents about resources that can help their children...

...PM--Catch up. Finish any communications with my UK team members before they log off for the day. 1:45 ...getting to talk to so many different people! I communicate with customers, software developers, as well ...from interacting with others, and have strong communication skills to tailor your message to any type of... to solve is clear, but the solution is more complicated. You’ll need to be creative to be able to work...team in the UK is five hours ahead, so I’m always catching up in the morning! 9:00 AM--Technical implementation...out. 4:00 PM--Respond to customer help tickets. Catch up on customer help tickets, and respond when I ...technology, software, and science. She loves to advocate for the customer and help build software and technology...

...guide outlines these components.     Fig. 1 - Viral vector components of retrovirus  Adeno-associated...ITRs serve as a type of origin of replication to initiate replication of the dsDNA virus. Viral packaging...plasmid while the other plasmids with packaging or replication components are called helper plasmids. Below,... production, they can be helpful for certain applications and aid in ease of use. Our lentiviral guide...AAV are rep and cap. Rep is required for viral replication and also aids in infrequent genome integration...a helper plasmid and mediate efficient viral replication. More information on these viral genes is available...particles. E2A/E2B are responsible for viral replication, with E2B being the primary viral polymerase....

...non-canonical amino acid at the site of interest (figure 1). While the codon used is often one of the three stop... its function, mimic a biologically relevant modification, or add in fluorescent or chemically reactive...function relationships or, in more practical applications, allow a protein to be more effectively used... you do to any number of experiments with. The catch The rapid expansion of GCE led to the creation of... of many new tools and potential applications. But this focus on development, along with the inherent ...’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques are straightforward...ligation through introducing ncAAs with protein modifications of interest or any of a huge variety of ncAAs...

...with each protein of interest.    Figure 1: The direct approach (left) versus the indirect approach... antibodies may be indicated on occasion, usually after other signal amplification techniques have been...IgG2 or IgG4. This can be important for some applications. Assay use While secondary antibodies are useful..., the flexibility of their use varies across applications. Western blots, for instance, almost always ...For the latter three, secondary antibodies are indicated when the protein of interest is found in low amounts...been ruled out.  Ease of access Signal amplification isn’t the only benefit of secondary antibodies. Finding...quantitative analysis of a sample. Instead, relative quantification can be done using either the direct or the ...

...prove the technology. This transition takes around 1 – 2 years. The product is the measure of success...competitions. After much learning, the focus of the application was changed, but Sample6 still leverages the ...fit for this set of technologies, effectively communicating the advantages embodied, and driving the enterprise...He explained that entrepreneurs must clearly communicate their idea at various levels of complexity and...backgrounds how amazing their idea is. While communication styles in academia and industry vary drastically...biggest challenge during this time was learning to communicate effectively to non-scientific audiences. "There...for the company; this will help avoid future complications and potential conflicts that waste money and...

...native peptide bond at the split site. Figure 1: The base editor is split into two plasmids and co-transfected...voltage indicator Article contributed by Shreya Vedantam Genetically encoded voltage indicators (GEVIs)...fluorescent optics. Existing red fluorescent voltage indicators, however, fall short compared to GFP-based sensors...linker between them. The Ace-mScarlet voltage indicator works like many FRET-opsin GEVIs. With an increase...channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019...increasingly been recognized for its biotechnological applications including conversion of CO2 into water and electrical...the N-terminus of Cre (CreN) are fused to the truncated Phytochrome B (PhyBΔ) and the active protein binding...

...give these stabilized promoters a try. References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag, and...rounds of replication are initiated before the cell has divided. This means that genes located near the...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...

...methods to improve AAV production.     References 1.Clark, K. Reed. "Recent advances in recombinant adeno-associated... Protocols for Work with Virus Watch Our AAV Purification Video ...

...regarding what makes this group of researchers special: 1. Stem cell scientists care deeply about ethics Ethics... have profound impacts on the development of applications with the potential to save lives. While most...

...-fDIO mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg...availability date or to be notified when a particular catalog item is available for order please email [email protected]...

... Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: 29551272 Plasmids from this paper ...for pre-clinical and clinical applications of dCasRx. The identification of Type VI Cas13d enzymes is ...RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader...even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable...target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding properties...

...sites that were previously unavailable. Figure 1: The PAM sequence that the Cas nuclease recognizes...to generate a genomic alteration at a specific location, it must be accessible by Cas9. Our previous blog...off-target activity, which may be undesirable in many applications. At the time of our first blog post on choosing...CRISPR-Cas9 to increase its specificity." Nature communications 9.1 (2018): 3048. PubMed PMID: 30082838. PubMed...CRISPR-Cas12b for human genome editing." Nature communications 10.1 (2019): 212. PubMed PMID: 30670702. PubMed...

...the output of the analysis performed in experiment 1. Use camelCase: In camelCase, each subsequent word..., specific goal for each analysis will help you locate relevant information in the future. Approach:...folders and files that you see on your desktop. The location of your master directory or folder for all of ...simple .txt document. Record the name, date, and location of all analyses (see above). Give every experiment.... She is particularly interested in science communication and epigenetics. Follow her on twitter @kc_palozola...

...speed: base editing before Cas lets go Figure 1: David Liu gives a talk at the Cell Symposia: Gene... 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...

...accessible to practically everyone.     Fig. 1: Open source hardware builds   Since I started...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...

...in Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while...INTEGRATE consists of four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade...for successful modifications, efficiency can be low, especially for larger modifications. What’s the solution...several potential CRISPR applications. First, CRISPR can be used to target locations in the genomes of pathogenic...tools available for bacteria and some of the applications for which they’ve been used. The beginnings ... of Enterobacteriaceae, without the need for modification. This finding suggests that the system may be...eukaryotes (Qi et al., 2013). As in other systems, catalytically dead (dCas9) targeted to a promoter or gene ...

...region: IgM, IgD, IgG, IgE, and IgA.    Figure 1: Diagram of an antibody labeled with Fc, Fab, heavy... Isotype controls are used in antibody-based applications including flow cytometry, immunohistochemistry...of these five isotypes, such as the number and location of disulfide bonds, the number of constant domains...

...of interest and the metal ions.      Fig. 1: A sample containing a polyhistidine tagged protein...proteins within a system. This can be pretty complicated given that a single cell has tens of thousands...single protein? Oftentimes, they rely on protein purification. In this strategy a single protein can be isolated...this blog will focus on one of the most common, purification based on the polyhistidine tag. What is the ...purity (Janknecht 1991, Hochuli 1988). In this purification approach, the metal ions are immobilized on ... incorporation of a cleavage site and a post-purification cleavage step to remove the tag in order to ...2000).  One downside of polyhistidine tag-based purification, however, is the propensity for nonspecific ...

...for RaPID is an RNA motif of interest. See figure 1 below for details on the key components of RaPID. ...methods Not in a native context. While native purification preserves RNA structure and naturally occurring...used the Contamination Repository for Affinity Purification (CRAPome), to identify highly biotinylated background...research groups that performed BirA*-based affinity purification mass spec experiments. These background proteins...subtracted from Ramanathan et al.’s RaPID datasets. Applications of RaPID RaPID sounds like a nifty technique...help answer? Ramanathan et al. outline three applications of RaPID: Discovery. To discover novel RNA-... that QKI is a host protein critical to Zika replication. Human genetic disease. RaPID can also be used...

... trafficking of these biosensors.   Figure 1: Schematic of iGluSnFR3 (left). GluSnFR3 in pMinDisplay... Haoyi Qiu He Tian Every two years, we seek applications for awards to early career scientists using ...virtual conference attendance. We also opened up applications to research technicians and undergraduate researchers...mercaptopyruvate sulfurtransferase (MPST) which catalyzes the desulfuration of 3-mercaptopyruvate and generates...the chromophore that are susceptible to redox modification, the MPST hydrosulfide can be transferred to... and CNO silencing. (D) Immunohistochemistry verification of the dual virus expression in CR neurons of...where she engineers genetically encoded voltage indicators. Tian developed a video-based high-throughput...

... an entirely new affinity reagent.    Fig. 1: A comparison of an antibody (left) to several alternative...re used in a number of research and clinical applications. Most of the time, when someone speaks about...deeper penetration into a tissue, useful for applications like IHC, or be able to access an epitope folded...that would size-exclude an antibody. In imaging applications, smaller affinity reagents means higher resolution...antibody’s ‘Y’. They are often used in clinical applications. Fab fragments can be created in lab in situations...Shahied, 2004). They are often used for clinical applications, including imaging.  Diabodies are ~60 kDa proteins...antibodies are still an excellent option for many applications, and frankly, if ain't broke, don't fix it!  ...

...see how you use them in your research!  References 1.Tang, Jonathan C.Y., et al. “A nanobody-based system...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...PubMed PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer, A., et al. “Modulation...

...many of which are available in rAAVs!   Figure 1: Diagram of AAV components      Cargo capacity...experiments. Recombinant AAVs (rAAVs) The primary application of AAV is delivery of genetic cargo to a host...numerical designations, separated by a slash, that indicate the ITR and serotype, respectively. For example...transduced gene is episomal and does not divide and replicate in the same way the host DNA does. Because of ... since you don’t have to wait for the host to replicate the DNA before expression. Now that you’ve got... S., et al. “Various AAV Serotypes and Their Applications in Gene Therapy: An Overview.” Cells, 12(5),...

...antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness because they accidentally...compensation.    Working with antibodies can get a bit complicated, but hopefully this helps you successfully select...

...internalization of two different receptors- angiotensin II type 1 receptor and the β2 adrenoceptor. GPCRs in chemogenetics...constructed plasmids that can be used in peroxidase-catalyzed proximity labeling to quantifiably measure GPCR...In this technique, the receptor fused to APEX2 catalyzes the biotinylation of proteins near the GPCR. These...Multidimensional Tracking of GPCR Signaling via Peroxidase-Catalyzed Proximity Labeling. Cell 169:338-349.e11 . https...Kadowaki S, Haga T, Takaesu H, Mitaku S (2002) Identification of G protein-coupled receptor genes from the...

..., there are two main decisions to make: 1. Which modification? and 2. Which viral vector should be used...excitation or inhibition upon light (or drug) application.  Genetic manipulation can also reduce expression...recombinase on transgenic flox animals to introduce modifications at the DNA level in a temporally and locally... brain area, avoiding side effects from such modification in other areas. An alternative strategy is to...

...best tips beyond the basics to get you started. 1) Be visible and open to connections This is a site... example, I will often see a whole bunch of notifications “a postdoc from this Uni viewed your profile...look straight at the camera. Pro tip: Turn off notifications to your network of profile changes. If you are...

... Science Find the SEND plasmids    Figure 1: SEND combines an endogenous Gag homolog, cargo RNA...AsCas12f1. They found that this system, without any modifications, is effective in genome editing in both bacteria...therapeutically relevant gene targets and expand the application to the possible treatment of genetic disease....

...Set a goal for paper-reading, even starting out at 1 paper/week will be useful. Check out the hashtag #365papers...looking for a particular protocol or are trying to replicate an experiment, the methods section probably won...is science’s way of validating work prior to publication, it doesn’t mean that there are no mistakes or...

... to the use of multiple antibodies. Figure 1: Overview of using Sandwich ELISA to quantitatively...the most common type of competitive ELISA is a modification of the sandwich ELISA as described here, the...ELISA, the only difference being the additional amplification step, which helps to visualize low-abundance...

...conclusions.  Figure 1. Potential points where DNA errors can be introduced, indicated by exclamation marks... utilize an array of tools that rely on DNA amplification and manipulation to produce and edit plasmids...places where DNA is being created, such as PCR amplification or bacterial growth. Created with BioRender....through the QC pipeline to resolve any issues.  Catching and addressing QC issues Addgene’s QC team has...has developed a process for identifying, categorizing, and resolving a variety of QC issues. This process...discrepancies can be caught prior to distribution. Catching these errors prevents future experimental issues...during analysis. These include topics such as: locations in a plasmid that are less likely to affect function...

...supervisor, even if he or she is a difficult person: 1) Understand your supervisor’s expectations Every ... series.  “The only healthy communication style is assertive communication.” - Jim Rohn, Author, Entrepreneur...I heard about supervisors in all of the above categories. In addition, I also learned about two other ...Assertiveness? An assertive person is able to communicate their ideas confidently while also considering...of the most frequent sources of conflict is miscommunication. For example, you might misunderstand your...the wrong direction. The best way to avoid miscommunication is to follow up after every meeting with an...workplace, these strategies will help you to communicate with confidence earn respect from your supervisor...

...basis of many molecular biology tools! Figure 1: Common functions of natural plasmids. Created with...Shareck et al., 2004). Plasmid categorization is complex Plasmid classification has shifted over the years...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although...genes between bacteria can also have serious implications for public health. Bacteriocin plasmids: microbial...poisoning, the difference is virulence plasmids! This category includes genes that help bacteria invade hosts...years, and new classifications are still being created as whole-genome sequencing becomes more relevant (... common plasmid features, such as origins of replication and promoters, check out our other Plasmids 101...