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We narrowed to 885 results for: ANT

Showing: 451 - 480 of 885 results

  1. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    Published
    Jan. 31, 2019, 1:32 p.m.
    ...on the conversion of non-malignant forms of cancer to invasive and malignant forms and noticed that myoepithelial...stromal cell modifications, stromal cells are also constantly communicating with a tumor, proving once again...malignancy but also incite different organs to become malignant. Exosomes also contain miRNA (micro RNA) and other... case, exosomes may transport miRNA from drug-resistant cells to sensitive ones. Though this process is..., the microenvironment backstage play just as important of a role if not more.  Many thanks to our guest...

  2. 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...

  3. 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...

  4. 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...

  5. 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...

  6. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...number of important plasmid elements – promoters, origins of replication, protein tags, and antibiotic resistance...recombination at the loxP recognition sites. A key advantage of the system is that Cre acts independently of...potentially recombine. To account for this, alternate mutant versions of the loxP site have been created, which...lox2272 (GgATACtT), and lox511 (GtATACAT). These variant lox sites undergo recombination with other sites...not cross-compatible. Using different lox site variants allows Cre to catalyze more than one specific ...

  7. 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...

  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. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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 ...

  16. 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...

  17. 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  ...

  18. 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...

  19. 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...

  20. 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: ...

  21. 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 ...

  22. 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...

  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. 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...

  25. 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...

  26. 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....

  27. 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...

  28. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    Published
    Jan. 23, 2020, 2:45 p.m.
    ...recipe. It’s unrealistic to plant, grow, and harvest grains every time you want to bake a cake. Instead, ...development happening at Addgene under the hood. A significant portion of our development may not be visible...changing needs of the teams at Addgene. Another advantage of Scrum is that it provides transparency to everyone... been updated in several years) or they make significant changes that make it incompatible with our software...software from Python 2.7 to Python 3.5. There are significant changes to the language and it took us a total...

  29. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...targets for complement-based killing by anti-alphaGal antibodies. The VSV-G envelope protein is inactivated...different viral membranes, tropism, and advantages and disadvantages: Table 1 (Joglekar et al., 2017)  Table...its own, it needs to be pseudotyped. This takes advantage of the EnvA-TVA receptor-envelope system (Wickersham...2006) Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods 4:47–49 . https://doi.org...

  30. 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...
Showing: 451 - 480 of 885 results