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We narrowed to 922 results for: Por

Showing: 331 - 360 of 922 results

  1. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    ...around the world, the spike protein has had many opportunities to evolve. Several variants are now circulating...different spike protein variants that can be incorporated into pseudoviruses. The lab used these pseudoviruses...Addgene to better understand which human genes are important for SARS-CoV-2 entry into human lung epithelial...). In their search, they found mucins have an important role in viral entry, which are inhibitors of SARS-CoV...research through sharing reagents has never been so important.     References and resources References ...

  2. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...experiments are important for visualizing ncRNA localization and function, but directly incorporating very long...transcribed but do not encode proteins. They play important roles in many cellular and regulatory processes...complex and not yet fully understood, so it is important to carefully select the target locations of knock-in...nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription machinery of a target...

  3. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...expression levels, the terminator also plays an important role in RNA processing and contributes to variability...polyadenylation. The addition of the poly(A) tail is important for stability of the mRNA, protection from degradation...degradation, and is integral to the nuclear export and translation processes as well.   Figure 2... non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and/or use...the Mason–Pfizer monkey virus constitutive RNA transport element lead to enhanced HIV-1 Gag expression ...

  4. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ... I can support that.” That was interesting. If people could be against GMOs as food but support GMOs that... up for real: bank account, business cards, incorporation, laboratory lease, etc. I still flip to that...result in a new utility, was there a business opportunity out there for us? After about a month of discussion...color, but the experience is invaluable. This opportunity has given us the chance to develop the technology...

  5. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...builds on existing popular web services, currently Crispor and Crispresso, that adhere to the same principles...decided to adapt the popular guide design tool Crispor by Max Haeussler. With a few extra lines of code... to analyze  CRISPR repair outcomes, which is important for quality control. The only ones we found that...analysis because of its ease-of-use and comprehensive reports. With CrispyCrunch, you get the all information...coding region in a UTR, CrispyCrunch flips the two important base pairs of the PAM (NGG -> NCC, CCN -> GGN)...

  6. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...detector using a series of mirrors and filters. Importantly, conventional flow cytometry uses one detector...broad, detectors will usually only pick up a small portion of that spectra to use for compensation.   ... flow cytometry for a given experiment, it is important to consider how many parameters you want to use...conventional and spectral flow cytometry, it is important to consider the expression levels of the markers...two highly overlapping cellular markers, it is important to pair them with two fluorophores that have distinct...

  7. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...plasmids/year from 2017 to 2023. This growth is supported by new labs joining the Addgene CRISPR community...year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized by ...plasmids deposited by country; square size is proportional to number of plasmids deposited by each country...plasmids requested by country; square size is proportional to the number of plasmids requested by each ...Barrangou for his authorial contributions to, and support for, this and previous Addgene CRISPR publications...

  8. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...Characterization of the Pink and Blue Pigments of Pocilloporid and Acroporid Corals. The Biological Bulletin 189:288...chromoproteins and fluorescent proteins, it’s important to note that despite their distinction by name...interactions with cytosolic proteins. This step is important as non-specific binding to proteins can inhibit...improved expression in living cells. Scientific Reports, 9(1). https://doi.org/10.1038/s41598-019-48604...

  9. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...diseases are also underway and reported results are encouraging. Importantly, a significant amount of basic...that can be monitored non-invasively over time. Importantly, it is an immune privileged compartment, meaning... large scale manufacturing of viral vectors to support trials for prevalent diseases - to name a few. ...combined with AAV for in vivo gene editing, and reports suggest that the first clinical study combining...

  10. MXS Chaining

    Type
    Blog Post
    ...produce complex fluorescent reporter constructs. These fluorescent reporters can be used to detect structure...), the same cloning procedure can be used to incorporate each element into the final construct. This strategy...protein sequences to develop diverse fluorescent reporters is a useful application of high-throughput cloning...sequences), so enzyme choice is critical. May not support in-frame fusions of coding sequences Sladitschek...

  11. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...research uses ancestry groups rather than self-reported race, it highlights the myriad issues researchers...designed a tool that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral...default” genome looks like, which led to the incorporation of ancestral bias into genome-wide screens, ...complicating the issue is the long and ugly history of reporting false essentialist links between biology and ancestry...design of the experiment or dataset itself. It is important for scientists to not only be aware of the potential...

  12. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...pathway is ubiquitous, with successful recombination reported in all laboratory E. coli strains tested so far...use of this pathway as a cloning tool was first reported over 25 years ago (Douglas and Howard, 1991, Bubeck...cloning strain has failed so far. What is most important is the transformation efficiency of the strain...primers that bind at either side of the undesired portion, designed to amplify the rest of the vector sequence...exploiting bacterial in vivo assembly." Scientific reports 6 (2016): 27459. PubMed PMID: 27264908. PubMed ...

  13. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ... in a particular organism. Why is codon usage important? Translation and protein synthesis relies on transfer...deliver amino acids to the ribosome where they are incorporated into the growing polypeptide chain. One part...even been linked to some diseases. It has been reported that 5-10% of human genes contain a region where...(Mauro and Chappell 2014).  Ultimately, it is important to take a close look at your experiment and gene...EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001 Additional resources on the Addgene blog...

  14. Bioinformatics at Addgene

    Type
    Blog Post
    ...this two-part blog series, we will describe the importance of bioinformatics at Addgene and how our search...Customizable to our specific needs Modern and well-supported, so we could focus on our development instead ...encouraged to see that a community of Nextflow supporters existed who had created a shared resource of ...modern architecture of Nextflow included built-in support for containers, such as Docker. If you’ve heard...other for the entire package to run. The small, portable system provided by a container is sequestered ...

  15. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...Their porous surface provides a large area for binding, but antibodies conjugated within the pores are ...used for IP. These proteins bind only to the Fc portion of the immunoglobulin, leaving the antigen binding...or a magnet (magnetic beads), providing a solid support while unbound proteins are washed away.  While ...to be adjusted depending on the sample. It is important to remember that sample lysates will also contain...

  16. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...either fold to incorporate their target molecule, such as a metal ion, or fold to incorporate themselves ...diagnosis. As biosensors, aptamers hold a few important advantages over their amino acid-based counterparts...highly customizable in terms of how they actually “report” as biosensors. Making aptamers glow The flexibility...ease of production have also made aptamers an important component of high throughput assays with microarrays...

  17. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...gene disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag antibody ...Western Blot for Anti-V5 [IPI-SV5-Pk1]. Addgene Report, https://doi.org/10.57733/addgene.l0ulu4. Hanke... expressed and purified from E. coli, retains important functional properties, and is amenable to a wide...doi.org/10.1128/jb.00335-23.  Bioluminescent reporting of mechanical forces with PILATeS By Emily P. ... reaction (Figure 5). The researchers used the reporter to visualize biologically relevant forces at focal...

  18. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...into cells by electroporation. hiPSC colonies are obtained by 10 days after electroporation, most of which...over a decade back when Yamanaka and colleagues reported that it is possible to reprogram differentiated...generation of human iPSCs (hiPSCs) now provides an opportunity to develop and use patient-specific somatic cells... blood, a huge improvement from the previously reported efforts. The pCXLE episomal system thus provides...

  19. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...and many other countries, and it is still imported, exported, and used throughout the world. New mesothelioma...being to cure cancer, more funding and research opportunities are being provided to immunotherapy than ever...that asbestos exposure remains the single-most important factor in developing mesothelioma. Asbestos, and...cells that can begin an immune response. This important biomarker can be used to distinguish between three...

  20. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ...a company to collaborate whenever it faces an important challenge. The types of projects mentioned above...intricate task of designing a strategic vision for a portfolio of candidate drugs. A decision like that would...scientific knowledge behind you, you’ll have the opportunity to learn the business side of consulting cases...or who needs a stepping-stone to many other opportunities. Many thanks for our guest blogger, Gairik ... Resources on Addgene.org Check Out Career Opportunities at Addgene ...

  21. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...that generates the chromophore (Tsien, 1998). Importantly, this reaction requires molecular oxygen and ...non-natural fluorescent amino acids and their incorporation into proteins are steps in that direction (Chatterjee...interest with an additional protein module that can report on location. This approach enables the study of...limitations. Being aware of the limitations is important for the successful use of GFP or any of the other...9759496.  3. Buckley, Anthony M., et al. "LOV-based reporters for fluorescence imaging." Current opinion in ...

  22. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...this article, we’ll go through some of the most important considerations to make when planning and performing...same sample. For this strategy to work, it is important to ensure that you are using secondaries with ...primary, so we’ll start there. Some of the most important considerations are to make sure your secondary...publications or as a review on a company website. It is important to review the data and protocols available for...regardless of how “risky” they may be, it is always important to verify their performance in your hands and ...

  23. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...Massachusetts Nonprofit Day, which was the perfect opportunity to test shooting over Zoom, everyone’s new favorite...directing. Everyone’s schedules were busy, so it was important for me to let people record at their leisure. ... were still many restrictions in place, most importantly, masks.  Besides the muffled voices beneath masks...out of the way when it was blocking some more important lab work from being done. You can watch the first...things return to relative normalcy, I’ll be incorporating that mobile monitor stand into every shoot. ...

  24. Tips for Getting a Faculty Position

    Type
    Blog Post
    ..., documents need to minimize jargon. Equally importantly, you really are selling yourself. A sales pitch...anyone could relate to, that people would agree is important to understand. (1-2 sentences). For example: Distinguishing... lot of time to discuss and the questions are important. However, it's much easier to focus on the questions...version of my manual, I added new chapters to incorporate the experiences of the people that provided me...interviews. We didn't have Skype in 2004. I also incorporated example documents. This part has generated a...

  25. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...of carboxyl groups to organic acids to produce important cellular metabolites. Biotin acts as a carboxyl...primary antibody respectively. Avidin bound to a reporter is then added to the reaction for easy detection... experiments. During detection, a biotinylated reporter enzyme that is pre-incubated with free avidin ...protein of interest leading to amplification of the reporter signal and easier detection. This strategy is ...biotinylated primer. Biotin can also be enzymatically incorporated during PCR using a biotin labelled dUTP. There...

  26. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...regulatory elements (to mutate promoters/enhancers in reporter constructs), in functional analysis of proteins... by an increase in the size of the lower band (portion of the plasmid containing the mutation). Similar...which could potentially truncate the template). Importantly, you also need a DNA polymerase that produces...resulting construct. In cases where the functionally important regions span several kb (or are GC-rich and therefore...you can simply amplify and mutate this smaller portion of the plasmid with a primer pair that generates...

  27. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... This is important to ensure that you’re looking at institutes that have the means to support your work...screening out irrelevant candidates, and, more importantly, it’ll help you define salary costs. 4. Ascribe...make these decisions in the following order of importance: 1. First make decisions based on experimental...and availability of the equipment is of utmost importance. Conducting a successful lab start up negotiation...

  28. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...methylase mutation (GATC) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes... mutation (CCWGG) Preparing unmethylated DNA, important when trying to cut with certain restriction enzymes...Allows uptake of large plasmids hee   High electroporation efficiency supE44 (glnV44)   Suppression ...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F-mcrA Δ(mrr-hsdRMS-mcrBC...propagating plasmids expressing the ccdB gene (important in Gateway cloning). F- gyrA462 endA1 glnV44 Δ...

  29. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ... homology arms and a selection marker, then electroporate it into mouse embryonic stem cells. After selecting...compatible with standard microinjection and electroporation methods using in mouse transgenics. Designing...between the ampulla and infundibulum. In vivo electroporation is performed immediately after injection. Image...tweezer-type electrodes are placed to mediate electroporation. Ohtsuka et al. compared traditional microinjection..., et al. "One-step generation of mice carrying reporter and conditional alleles by CRISPR/Cas-mediated...

  30. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...injection site  (Kanaan et al., 2017), axonal transport  (Murlidharan et al., 2014), and postsynaptic ... system, remember that some serotypes can be transported retrogradely and/or anterogradely along the axon...et al., 2014; Nieuwenhuis et al., 2021). It is important to note that this expression pattern can be affected...of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse...Favre, D., Blouin, V., Provost, N., Spisek, R., Porrot, F., Bohl, D., Marmé, F., Chérel, Y., Salvetti,...
Showing: 331 - 360 of 922 results