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Showing: 451 - 480 of 885 results

  1. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...Accelerate Research Discoveries) Challenge provides grants for rare disease research. This year is our fourth... 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...genes from the X chromosome are expressed at significantly reduced levels in women with TS (Trolle et al...nucleus,” says Dr. Terry Hébert, a professor and assistant dean of biomedical science education at McGill.... So far, there have been thirty GNB1 missense variants identified from 58 individuals with GNB1 disorder...

  2. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ... nuclease-resistant, resulting in less off-target events. While both have similar advantages, bridged ...Nature has evolved ways to prevent degradation of important cellular RNA molecules with post translational...-Cas9, for use in applications like siRNA and antisense oligonucleotides. Some of these synthetic modifications...Frontiers in Genome Editing). This is particularly important in experiments in which viability is being compromised...technology is that most expression systems are constantly “on”. There is no mechanism to turn cutting off...

  3. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    .... 1). When you have to tell an engineer that you want the LIMS software to create a barcode for your plate...each given container, when applicable Any other relevant information Prescribing an official name to all... all of our plates, tubes, and racks meant we had fewer errors in our LIMS tracking system. Though you...terminology of a new field. Annotate: When creating important documentation, including SOPs or project-related...Product Manager role at Addgene is “Words are important!”  If you try any of these approaches in your ...propensity for cross team communication, that’s an important soft skill to carry into the next phase of your...

  4. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    Published
    Nov. 3, 2020, 2:15 p.m.
    ...high-intensity light for activation which requires implantation of fiber-optic cables into the brain. ...information about parental channelrhodopsins to create recombinant channelrhodopsin sequences. The model first removed...performed well in a tissue culture dish, that doesn’t guarantee their success in vivo. Therefore, the Arnold and...fiber-optic cannulas were still stereotaxically implanted into the mice’s brains so that light could reach...into a hole in their cage. Since dopamine is an important neurotransmitter that promotes reward-motivated... brain may still require fiber-optic cannula implantation, but altering ChRgers so they can be activated...

  5. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...transcription factors with important roles in Multiple Myeloma), it is important to ensure lack of degradation...Remarkably, small molecule degraders provide distinct advantages over pharmacological inhibitors including rapidly... are happy to share aliquots of dTAG-13 in the meantime, please address your requests for dTAG molecules... chimeras For each protein-of-interest, it is important to determine which terminus can accommodate the...context. Immunoblotting is recommended, while quantitative proteomics experiments will provide the best...

  6. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ... and Timothy K. Lu. "Enhanced killing of antibiotic-resistant bacteria enabled by massively parallel combinatorial...particular phenotype, and quickly profile the resultant hits (Cheng et al., 2014; Wong et al., 2015; Wong...plasmid. Each gRNA combination can be tracked and quantitatively analyzed by sequencing its set of barcodes....efforts made by various research teams to build and constantly improve gRNA libraries, excellent resources with... Wong et al 2015. Alan Wong is currently an Assistant Professor at the School of Biomedical Sciences...

  7. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...expression, plant expression, mouse targeting, CRISPR, TALEN, synthetic biology, affinity reagent/antibody, unspecified...be indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as ...other. Bacterial Resistance Enter bacterial antibiotic resistance encoded in the plasmid that should...or low copy. Choose “high copy” if a sufficient quantity of DNA is produced from a miniprep and “low copy...Pro Tip! Download a new spreadsheet each time you want to submit new plasmids! We update the spreadsheet...

  8. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...updated our blog post on Prime Editing, and that meant rereading many of the original papers reporting ...editing application. And optimizing its sequence is important, as prime editing efficiency is affected by the...nucleotides. For longer templates, it is even more important to test different lengths and sequences, as unintended...whose 3′ extension begins with a C base can cause unwanted base pairing (left), while pegRNAs that use other... Tool (pegLIT) to create linkers that minimize unwanted intra-RNA base pairing with the primer binding...prime editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652...

  9. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...weeks designing and cloning the vectors you actually want. But now you can use the recently developed third...website, users can simply select the components they want for each module, generate the vector file, and quickly... license.   Once you have the components you want, simply assemble them into a destination vector ...vectors are available? What you’re building is important — but so is where you’re building it! Table 1 ...selections) Module set (the set of components you want in your vectors) Target type (drop down options...kit. What if Fragmid doesn’t have the components I want? You can make your own Fragmid components! Ensure...

  10. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ... series). In any case, Cre is a dominant phenotype, while you’ll want the floxed gene to be homozygous...these strains should be crossed to generate double-mutant offspring. Crossing mice allows for the production...homozygous for one of the two mutations that you want combined in your offspring. Genotyping involves ...generations. 4. Continue breeding your colony You’ll want to generate enough mice that are homozygous (or ...require several rounds of breeding in which you’ll want to genotype your mice for expression of Cre, loxP...

  11. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP variant, mGold. mGold... were exposed to blue light. Cre-Vvd showed substantially improved activity as compared to other recombinase...CRISPR-Casɸ is active in vitro as well as in human and plant cells. Casɸ from Biggiephage can eliminate...Birsoy. Find the library here A new SaCas9 ABE variant (microABEI744) has improved on-target editing efficiency...compared to current N-terminal linked SaCas9 ABE variants. It's also one of the smallest AAV-deliverable...

  12. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ... of the fusion protein. Advantages of Split-BioID Split-BioID’s key advantage over other PPI methods, ... lysate fractionation. Important considerations for Split-BioID It’s important to remember that with Split-BioID...proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that it allows for the validation...bait protein is pulled down, typically with an antibody. This can artificially select against PPIs that...

  13. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    Published
    May 22, 2015, 4:58 p.m.
    ...vector that encodes both Cas9 and sgRNA. What if I want to work with gene drives? Then do so! It's a tremendously...that anyone else who came up with the same idea or wanted to build a gene drive in a model organism would...experiments2,3. While that didn't work quite as well as anticipated4, greater awareness will certainly improve safety...regions that don't have resident populations of the relevant mosquito species. Reproductive confinement involves... cas9 gene). An immunizing reversal drive is a variant that also spreads through the wild population and...gene drives. Science 345, 626–628 (2014). PubMed. Gantz, V. M. & Bier, E. The mutagenic chain reaction: ...

  14. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...identical molecules of DNA. Gene targeting takes advantage of this natural process to replace a targeted ...marker HSV-tk is used to select against random recombinants.   Design your targeting construct. For...been cloned into the vector on either side of an antibiotic resistance gene. To avoid selecting for cells...will first perform a positive selection for the antibiotic resistance gene and then a counter selection ...Remove the positive selection marker. Since an antibiotic resistance gene can affect a cell's phenotype...a true knockout can be lethal, and you'd instead want to create a conditional knockout. To make a conditional...

  15. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ... to achieve: Do you want to permanently knock out or knock-in a gene? Do you want to activate or repress...trying to create a single point mutation? Do you want to create a fusion with a reporter protein such ...sufficient? Which cell types will you be editing? Do you want to deliver the components as DNA, or would mRNA ...and Cas9 are required for any application. If you want to include a homology directed repair (HDR) template...well characterized in terms of CRISPR use, you will want to invest some time in optimizing and testing the...

  16. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ... expressed proteins. Here we will review what advantages degron tags can offer over other protein control... protein expression. These tags offer several advantages over the alternative systems – CRISPR, siRNA,...function, the SMASh tag self-cleaves itself when you want the protein around, leaving a minimal additional...with the most alternative functions – there are antibodies to it and ways to achieve fluorescence with it...might be the way to go (dTAG/SMASh). If you don’t want to deal with additional components beyond the minimum...Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...

  17. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ... cells. To study this phenomenon you will likely want to monitor the voltage change across the membrane...comparable to its one component counterpart and is significantly brighter and available with red-shifted fluorophores...for penetrating deeper tissues depths (1 mm), important for certain applications such as in vivo tissue...encoded component of the system to offer unique advantages depending on your experimental system.   ...and this voltage is particularly physiologically relevant in neurons, myocytes, and endocrine cells. It ...sensor selectively is especially essential. If you want to learn more about how viruses can be used for ...

  18. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...chromoproteins and fluorescent proteins, it’s important to note that despite their distinction by name...photosynthetically active wavelengths to reach zooxanthellae, the coral’s symbiotic dinoflagellates that ...be used for mimicry. Image: Bernard Spragg. Advantages of using chromoproteins for molecular biology...bacteria, where most genetic manipulations start. Anthony Forster’s lab used codon optimization programs ...interactions with cytosolic proteins. This step is important as non-specific binding to proteins can inhibit...Characterization of a Blue Chromoprotein and Its Yellow Mutant from the Sea AnemoneCnidopus Japonicus. Journal...

  19. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ...when the hood is in use. Wipe the hood with a disinfectant (most people use 70% ethanol, but you can also...wipes) before and after your work. Keep your contaminants away While it’s likely that you’re the main ...pipettes into solution bottles directly, rather decant the solutions into disposable sterile tubes first...already be at 37 °C. Most labs have a water bath constantly at 37°C for this purpose.Learn about the effects...microscope will help you identify bacterial and yeast contaminants. Set up a routine to split cells Don’t be lazy...passage are too young and should not be used for important transfection experiments right after they’ve been...

  20. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... between the microhomologies.MMEJ has certain advantages over HDR. Some species don’t have good HDR systems...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus...into the FBL locus. Upon sequencing of puromycin resistant clones, they found that 80% and 50% of clones ...technique can be adapted to other transgenes. It’s important to note that the fluorescence level observed will...follows. For increased versatility, it would be advantageous to adapt PITCh to insert genes into AAVS1, the...to insert cDNA constructs into this locus. One important question to ask is how the efficiency of PITCh-based...

  21. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...systems. Can you describe this technology and its advantages? Fujii: In the last several years, my lab has...is being involved in the scientific community important to you and your lab? Fujii: I first knew Addgene...considering the peer-review system of journals and grant applications, for example. However, at the same ...interact with each other, which is critically important, especially for young researchers. It was a really...living for Japanese, which is not trivial, and the grant situation is currently much better than that in ...tough competition for academic positions and poor grant situation. So, young researchers need to publish...

  22. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...). Transformed bacteria are then grown up on antibiotics such that only bacteria with the plasmid can ...filtered pipet tips and prepare fresh media with antibiotics at each stage. The barcodes and LIMS help us ...enter the cells. Some samples require a reduced antibiotic concentration or need to be transformed into ...tubes. 1D barcodes are better for tubes that are meant to be read one at a time. 1D barcodes can be purchased...misread.  2. Track the container - It is just as important to track the location of the container housing...to fulfill orders.  The barcoded tubes help to guarantee that no samples get lost during their journey....

  23. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...Addgene's Antibody Guide Addgene's Ready-to-Use Antibody collection Addgene's Recombinant Antibody Plasmids...sure that the sample is pure and free of any contaminants that could affect the measurements. You'll need...below 0.95! More resources on the Addgene blog Antibodies 101: ELISAs The Four ELISAs and When to Use Them...

  24. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    Published
    July 9, 2019, 12:52 p.m.
    ...think of themselves as “scientists.” We had some accountants running our math tables!  Be realistic about...exposure. If you expect a lot of kids, you’re going to want a lot of activities (try for a dozen!). The best...presentation because, in my opinion, it was more important to communicate the applications of CRISPR over...use it? This last section was arguably the most important, and it was a little worrisome for me since bioethics... next year. There are many reasons why you might want to reach out to your school community as a scientist...scientist. Perhaps you have young children and you want to make sure their STEM education is supplemented...

  25. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ... likely seem important for any virus, we will see that they are very central determinants of how AdVs ...toward saying that host-virus interactions are an important part of the biology of adenoviruses (AdVs). AdVs...from the genomic plasmid can be used to house a recombinant sequence (transgene) instead.    Although the...natural AdVs, replication-competent viruses with a recombinant genome, and adenoviral vectors all have a genomic...basic research, like other viral vectors, though significant challenges remain (Wold & Toth, 2013).  While...

  26. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...the cells’ germline genetic variations. They then wanted to identify cancer-associated genetic dependencies...ancestry, Sean and Aaron knew that their study had significant statistical limitations, and expected to find...Cas9 from cutting, both of which can introduce significant bias into the results. But the reference genome...design of the experiment or dataset itself. It is important for scientists to not only be aware of the potential...ancestral bias and the human genome Wang, T., Antonacci-Fulton, L., Howe, K. et al. The Human Pangenome...doi.org/10.1038/s41586-022-04601-8 Lewis, Anna CF, Santiago J. Molina, Paul S. Appelbaum, Bege Dauda, Anna...

  27. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    Published
    July 1, 2022, 1:15 p.m.
    ...choose? Which way is the right way?  First of all, I want to put to rest a common misconception. I find personal...natural - if it works so well for you, of course you want to share it with others! After all, won't it work... to where your audience already is.  You’ll also want to keep in mind that access and availability to ...resources do you have?  No matter the project, I can guarantee you that either resource or time restrictions ... design a hands-on interactive activity using elephants than it is to design one demonstrating mitosis...Consider also the amount and type of information you want to convey. Small tidbits and fast/fun facts are ...

  28. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ... maybe you’re researching a topic for which no relevant or recent review exists. Once you pick a topic...format also has the benefit of exposing you to relevant researchers in the field. Finally, you can sound...from start to finish. Find the information that’s relevant to you, note it, and move on. At the beginning...retreat to the woods for months at a time, so take advantage of whatever time you have. If you’re like me, ...were, and you could find the original paper if you wanted to learn more. Another example: we made a schematic...Tips for Improving Your Next Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out Our...

  29. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...between the purine bases (adenine, guanine, and hypoxanthine) or between the pyrimidine bases (cytosine, ... purine to a pyrimidine or vice-versa. Note: hypoxanthine (shown) is the nucleobase component of the nucleoside...nucleoside inosine consists of the nucleobase hypoxanthine + a ribose sugar. Their fusion of wild-type ...glycosylase / alkyladenine DNA glycosylase  Removes hypoxanthine from DNA, creating an abasic site. The same ...one name. Indel Insertion/deletion A type of unwanted DNA edit. AYBE, AXBE, ACBE Adenine transversion...models-assisted optimization of a uracil-N-glycosylase variant enables programmable T-to-G and T-to-C base editing...

  30. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ... L., Martínez-Antonio, A., Quiñones-Valles, C., Espíndola-Serna, L., & Martínez-Antonio, A. (2011). Mechanisms...base pairs for the protein to be translated. An antisense RNA called Inc, also encoded by the ColIb-P9 plasmid... the host with a beneficial function (usually antibiotic resistance).  In the lab, low copy plasmids may... competition between pseudoknot formation and antisense RNA binding at a specific RNA site. The EMBO Journal...Leonard, A. C. (2018). Origin recognition is the predominant role for DnaA-ATP in initiation of chromosome...
Showing: 451 - 480 of 885 results