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We narrowed to 727 results for: cat.1

Showing: 601 - 630 of 727 results

  1. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ...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...

  2. Sharing is Caring: But How to Distribute Open Hardware?

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    Blog Post
    ...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...

  3. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...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 ...

  4. Antibodies 101: Isotypes

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    Blog Post
    ...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...

  5. Giving gRNAs a Facelift - Synthetic and Beyond

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    Blog Post
    ...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...

  6. To Each HIS Own

    Type
    Blog Post
    ...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 ...

  7. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ... 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...

  8. RaPID Detection of RNA-protein Interactions

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    Blog Post
    ...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...

  9. Antibodies 101: Affinity Reagents

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    Blog Post
    ... 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!  ...

  10. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...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...

  11. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...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),...

  12. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ...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...

  13. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...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...

  14. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

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    Blog Post
    ..., 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...

  15. What's New in CRISPR - November 2021

    Type
    Blog Post
    ... 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....

  16. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...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...

  17. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

    Type
    Blog Post
    ...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...

  18. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ... 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...

  19. A Look at Addgene's QC Process

    Type
    Blog Post
    ...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...

  20. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

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    Blog Post
    ...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...

  21. Plasmids 101: The Wide World of Natural Plasmids

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    Blog Post
    ...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...

  22. Illuminating Epigenetics with A FRET Based Biosensor

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    Blog Post
    ...treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon...Mechanisms of epigenetic modifications The main mechanism of epigenetic modification is chromatin remodeling...literally means ‘on top of the genome’). Epigenetic modifications can result in phenotypic changes without any...any change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling...two main mechanisms, either through histone modifications, which can influence the transcription of genes...hypomethylation, on the other hand, has also been implicated in the development and progression of cancer ...enzyme digestion, DNA adenine methyltransferase identification, and bisulfite sequencing. However none of ...

  23. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...If you already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise ...well as suitable applications. Expression System Components of System Application Mammalian expression...CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods.   ...Select the appropriate CRISPR tools for your application: Wildtype Cas9 or Cas9 nickase are appropriate... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator...or more sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair...Cas9-gRNA complex and delivered to cells using cationic lipids. Transient expression of CRISPR components...

  24. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...development programs for bioscience educators. Using these hands-on tools, educators introduce their students to...biologists have always struggled to effectively communicate the nuances of protein structure in a flat 2-...dangerous. Subscribe to CRISPR Blog Posts How do educators and students use molecular models?   We created...something tangible. Under the guidance of a trained educator, these instances of focused attention become magical... their labs. Soon after we began working with educators in our Center for BioMolecular Modeling, we spun...through flexibility It is common knowledge in the education community that All models are wrong….Some models...

  25. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...viruses - 2 cas systems regulate at the DNA level and 1 at the RNA level. Udi Qimron from Tel Aviv University...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...Guangzhou and the Science City The conference was located in the modern, relatively new “Science City” area...and technology companies and start-ups choose to locate there. I made time to go into the city of Guangzhou...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

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

    Type
    Blog Post
    ...which you’ve already fluorescently labeled. Figure 1: Using Split-BioID to Identify Protein Protein Interactions...methods for identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability ...the validation of a binary PPI as well as the identification of additional interacting factors. Problems...detect weak and/or transient PPIs. Many PPI identification techniques rely on protein interactors to stay... AP-MS. This requirement often prevents the identification of weak and/or transient interacting proteins...spatiotemporally defined protein complexes. Nature communications. PubMed PMID: 28585547. PubMed Central PMCID...

  27. Hot Plasmids - October 2020

    Type
    Blog Post
    ... inhibitory synapses in the brain.  Figure 1:Schematic diagram of different FingR constructs and... labeling proteins with different subcellular locations. Image from Goudeau et al., 2020. Goudeau... show a high proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in ...in high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...Hybrid for Multiplexed Editing and screening Applications (CHyMErA) Knockout Libraries express hybrid ...

  28. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...is best for larger genes such as Cas9. See figure 1 for an example of how changing transfection ratios...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct... as resources available, cost and downstream applications. For example, while the PEG precipitation method...

  29. CRISPR in the Clinic

    Type
    Blog Post
    ...significant progress in clinical trials.     Figure 1: Types of delivery and editing strategies for CRISPR... therapeutic potential in mice with a dominant cataract disorder caused by a single copy of a gene known...known as Crygc (Wu et al). They found that the cataracts phenotype could be rescued by zygote co-injection...substantial relief from both disorders. As of publication, submissions to the UK, EU, and USA federal drug...10.1186/s12943-022-01559-z Zarghamian P, Klermund J, Cathomen T. Clinical genome editing to treat sickle cell...

  30. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-...methodologies, with a special emphasis on these five categories. Please note that many of the following cloning...relies on site-specific recombination reactions catalyzed by BP Clonase and LR Clonase enzyme mixes. After...there is no need for restriction digests, gel purifications, or ligations. The general Gateway cloning protocol...
Showing: 601 - 630 of 727 results