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We narrowed to 847 results for: CAL;

Showing: 341 - 360 of 847 results

  1. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology...has facilitated the development of a simple and scalable means of making targeted genetic edits. Cas9 is... a gene of interest, they have been shown to dramatically alter levels of expression. Cas9’s ability to...Lance-Byrne is a graduate student at the University of California, Santa Cruz, where her research is focused on...Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease." Nature biotechnology ...Options for CRISPR Genotyping Cas9 Activators: A Practical Guide Additional Resources on Addgene.org Check...

  2. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ... editing. Specifically, these types of CRISPR systems work using a multi-Cas complex called Cascade (CRISPR-associated... Cas3. Cas3 can induce genomic alterations on a scale that other Cas effectors can’t, while maintaining...recruit Cas3, which carries out the brunt of the mechanical work for editing. Class 1 CRISPR systems make...unraveling the double-stranded DNA (dsDNA) in what is called a reeling mechanism (see next section for details...debilitating disease! This is, of course, only hypothetical at this point. More time and energy must be ...

  3. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ...protein must be expressed (inducibly, ectopically, stably… it’s your call). The TIR1 F-box protein will associate...their fused protein under certain conditions (typically the addition of an exogenous small molecule). ... size from several kDas up to ~35 kDa and are typically knocked-in via gene targeting or cloned in-frame...proteasomal degradation. Adapted from Nabet et al., Chemical Biology, 2018.    HaloTag The HaloTag was ...protein structures, interacting partners, and localization. Thankfully, this is a hot topic with many resources...

  4. Of Myc and Men

    Type
    Blog Post
    Published
    Jan. 19, 2023, 2:15 p.m.
    ...antibodies for every potential new target and basically impossible if you wanted to distinguish between...C-terminus post-translationally, which turned out to be critical for antibody recognition. As such, the peptide...interest and proteins tagged with it needed to be chemically treated before the antibody could detect them...investigating mechanisms of endoplasmic reticulum localization, Munro and Pelham casually introduced the Myc-tag... recognized by its antibody without additional chemical modification steps and when the epitope was within... of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable regions ...

  5. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ... the pharmaceutical industry for years. They have proven very difficult to target therapeutically due ...however, have yet to slay the beast. On a global scale, tens of millions of new cancer cases arise and ...fit. Kevan Shokat’s group at the University of California, San Francisco, is approaching the Ras problem... cancers, and synthesizing compounds to bind specifically to their mutated residues. For Ras researchers...represented, making the results of any such study as clinically relevant as possible. Connecting the research...

  6. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...delivery methods below: Chemical transfection   Lipid mediated Cationic polymers Calcium phosphate Cells... some CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator... be constitutive or inducible. U6 promoter is typically used for gRNA. May contain reporter gene (e.g....Medium Medium Medium Difficulty Easy Easy Easy Physical transfection   Electroporation / Nucleofection...

  7. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...is certainly not the first means to carry out so-called “forward genetic screening experiments”, it is ...in a particular phenotype. CRISPR can be readily scaled up for genome-wide screening due to the broad range...experiments you use them for! A well-developed biological question and experimental system are absolutely... to skewed results. Selecting a cell type - Theoretically, any cell type can be used in a CRISPR screen...screens check out Addgene’s 2016 publication “Practical Considerations for Using Pooled Lentiviral CRISPR...McDade JR, Waxmonsky NC, Swanson LE, Fan M (2016) Practical Considerations for Using Pooled Lentiviral CRISPR...

  8. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ... non-scientist friends and family is Part Five, called simply “Bad Science.” Zimmer kicks off the section...science, or misuse science, and that is what I would call bad science.”  The three chapters in this section...Zimmer provides several. In Part Three of the book, called simply “Old Science,” he describes the now infamous...are “old,” they lead Zimmer to one of his most critical observations: “Today the most successful scientists...optogenetics and CRISPR of course address some of the ethical implications of using these technologies, but also... certainly still is and will continue to be a “critical consequence of ...the marginalization of science...

  9. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...immunoprecipitation (enChIP) using CRISPR." Biochemical and biophysical research communications 439.1 (2013):...genomic alterations. These studies utilize a catalytically inactive form of Cas9 known as dCas9 (Jinek ...interactions at this site (Fujita and Fujii, 2013). They call this method engineered DNA-binding molecule-mediated...begun to make important discoveries in various biological fields. In 2015, the Fujii Lab identified non-coding... Fujita, Toshitsugu, et al. "Identification of physical interactions between genomic regions by enCh IP‐Seq...

  10. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...neuroscience research is the lack of a method for genetically manipulating neurons throughout the whole brain...CNS and PNS.That is until the Gradinaru Lab at Caltech stepped up to the challenge. Find the PHP Plasmids...astrocytes. Why screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but...throughout the body and is often difficult to surgically access. Therefore an IV injection is an ideal...pictures (see figure 4). It also enables single-cell anatomical study and neuronal tracing, aka determining where...mechanistic studies and gene therapy treatments for neurological diseases). The Gradinaru Lab has a lot going...

  11. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...  Ampicillin Ampicillin, often just called amp, is the historical king of the antibiotic resistance realm... antibiotic resistance genes so that you can specifically isolate bacteria containing your constructs ...immediately after it has been heat-shocked or electrically zapped. As a result, a shorter recovery period... of antibiotics, which induce cell death by intercalating into DNA and generating DNA breaks. The Sh ble... antimicrobial agents in MIC trays. Journal of Clinical Microbiology, 23(5), 959–961. https://doi.org/...

  12. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    Published
    July 1, 2025, 1:15 p.m.
    ...Thanks to the Beer–Lambert law, the instrument can calculate a sample’s nucleic acid concentration based on... reading, distorting the final concentration calculation and potentially impacting downstream experiments...in many plasmid purification kits), which can dramatically reduce the A260/230 ratio at concentrations ...systems to better identify contaminants and to calculate corrected concentrations. However, such systems...sound techniques, routine machine maintenance, and critical interpretation of the results.  If you see evidence...1ea8aec3bfa24543a28fcaea25986514.ashx. Koetsier G and Cantor E. A Practical Guide to Analyzing Nucleic Acid Concentration ...

  13. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...using a mammalian production system, cell lines typically used include Human Embryonic Kidney (HEK) cells...also need to know the extinction coefficient to calculate the concentration. At 280 nm, the extinction coefficient...protein standard and the concentration may be calculated using imaging software. Any bands on the gel ...validated to ensure they bind their antigens specifically. Possible applications for validation include...Sellick C, Varley P, Rockberg J, Chotteau V. Small-scale bioreactor supports high density HEK293 cell perfusion... MS, Carpenter JF, Lengsfeld CS. Do not drop: mechanical shock in vials causes cavitation, protein aggregation...

  14. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...the bacterial population. But what if we could specifically target only virulent or antibiotic resistant...antibiotics by their suppliers, and public health and medical organizations are working to help educate the public...encoded by a high-copy plasmid. In the case of the genomically encoded quinolone resistance, mediated by a single...represented here by cells of different colors, specifically targets bacteria with the given genomic or plasmid...Yosef et al. suggest that it could be valuable in medical settings where antibiotic resistant bacteria have...narrow bacterial host range. To further potential clinical applications, Ando et al. recently engineered ...

  15. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...scientists often refer to as “retroviruses” are technically gamma-retroviruses - another, separate member...differences may seem subtle they result in a key physiological difference between lenti- and retroviruses. ...transfer vectors for transient transfections? A2: Technically, yes. Will it actually work? That depends. The...when using a plasmid backbone that was designed specifically for transient expression (find some useful backbones...RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more than 30% of...terms of biosafety? A8: According to the American Biological Safety Association, "the two major risks of lentiviral...

  16. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...detection is crucial for clinical diagnostics, genotyping, and biotechnological advancements. Many methods...be used for an array of applications including clinical diagnostics (e.g. pathogen or virus detection)...points out that the protocol is not approved for clinical use at this point, they hope that the protocol...this ability to create a DNA detection platform called DETECTR. How does DETECTR work? DETECTR works similarly...mixed population for an array of molecular and clinical diagnostic applications. The company Mammoth Biosciences...

  17. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...improved, by adding a nuclear localization sequence (SV40 Large T nuclear localization sequence) and expressing...asymmetric target site core but its 8 bp length that is critical for Cre and FLP function. Because recombination...Recombination between the two target sites with identical sequence leads to inversion of the flanked DNA...this plasmid mix results in sparse labeling of cortical neurons and their synapses.   How can you...control   Figure 2: Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry...

  18. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...range of logistical barriers. Addgene worked to identify, reduce, and remove these logistical barriers...’ve turned the available into the accessible. Physical access Addgene is located in the USA, but our ...Beijing Zhongyuan 2012 China Summit Pharmaceuticals International 2013 Japan Leehyo Bioscience...Digest Analysis We also introduced guides — practical primers to help users get started with tools like...increasing the accessibility of our repository, from physical materials to useful information, but we’re not...

  19. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ...a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such as horseradish...level of antigen present in the sample and is calculated by extrapolating from the standard curve. Because...include undesirable molecules that may bind non-specifically to assay components and increase background ...amount of antigen present in the sample and is calculated by extrapolating from the standard curve. Scientists...antibodies are used.  Competitive ELISAs Scientists typically use a competitive or inhibition ELISA when measuring...While the precise setup of the method can vary, typically users first coat the multiwell plate with a reference...

  20. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...DnaA binds multiple specific sites within the ori called DnaA boxes. These boxes come in both high- and ...Created with BioRender.com. Once DnaA breaks the local hydrogen bonds holding the DNA together, it recruits...itself. RepA binds to directly repeated sequences called iterons located near the ori and directly interacts... the protein to be translated. An antisense RNA called Inc, also encoded by the ColIb-P9 plasmid, is complementary...between iterons on different plasmids in a process called "handcuffing," which blocks replication. ColIb-...Replication Browse other Plasmids 101 Posts Get practical molecular biology help with our Molecular Biology...
Showing: 341 - 360 of 847 results