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We narrowed to 910 results for: ire;

Showing: 541 - 570 of 910 results

  1. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair (HDR) template...straightforward. Likewise for step 4, as this ties directly back to your specific goal. When thinking about...

  2. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...dCas9, require Zinc-finger or TAL proteins targeting a specific genomic locus, which can require multiple...genomic region was an intensive process. One method required insertion of a recognition sequence into the genomic...

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

    Type
    Blog Post
    ...big science,” a massive, expensive project that required collaboration from scientists around the world... undermining science are even more immediate and dire than Zimmer could have imagined. Does this detract...about CRISPR alone, and Zimmer himself has written entire books about optogenetics. Defining science and ...

  4. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...to ddPCR. Droplet digital PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents...20,000 droplets. Each droplet contains the material required for a mini amplification reaction to take place...for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and Eriko Uchida. "2D droplet...

  5. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...mammalian antibiotic selection gene. However, it does require a slower transformation recovery period than amp...zeocin resistance by expressing a protein that directly binds to zeocin, preventing it from binding DNA...low rate, there is evidence that zeocin is never entirely detoxified, even when Sh ble is expressed (Trastoy...

  6. An Introduction to Adenovirus

    Type
    Blog Post
    ...sense of what AdVs are about – and we hope that it inspires further use of existing vectors, as well as research...response to adenoviral vector infected cells does not require de novo viral gene expression: Implications for...1996). The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient...

  7. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally...either briefly boiling the sample before use or by directly adding a small volume of the sample to the PCR...tell you whether the insert has the correct directionality. Verifying the insert sequence with Sanger ...

  8. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...from long written proposal narratives. Now they require grant seekers to use online applications with significant...to and proof of compliance with the grantor’s requirements as well as state and federal regulations. A ... worked for an organization in a position that required some involvement in the grants process and managing...

  9. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...generating reliable results from CRISPR edit data requires the help of robust software tools. As a consequence...of the edited sample. You can also download the entire analysis as a ZIP file by clicking the Download...the inferred distribution of indel sizes in the entire edited population of genomes. Hovering over each...

  10. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral production...efficiency can vary across cell types and generally requires optimization of many variables, such as seeding...Using pooled libraries can be intimidating and require a large amount of experimental work, but with this...

  12. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    .... Cre-dependent expression of the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive...neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly demonstrated...., 2018). The pooled nature of this experiment requires fluorescence activated cell sorting (FACS) to ...

  13. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...on pEcgRNA plasmid as only two 24 nt oligos are required and the BsaI linearized pEcgRNA backbone can be... in large amounts and frozen for future use or directly ligated with the annealed 24 nt oligos to generate...the above systems are great for experiments that require only single chromosomal integration (ex: genome...

  14. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ... sites. In cases like these, antibodies can be directly conjugated to agarose or magnetic beads with commercially... commercially available chemical agents. Directly conjugating the primary antibody has the additional ... particularly advantageous because they do not require centrifugation between washes which can cause loss...

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

    Type
    Blog Post
    ...recombinase recognizes FRT sites and initiates site directed gene recombination between those sites. FRT sites...with low concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors...PMC41457. 8. Schnütgen, Frank et al. 2003. “A Directional Strategy for Monitoring Cre-Mediated Recombination...

  16. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...exposed even one time. Doctors and researchers don’t entirely understand why some people develop the disease...one of the best ways to improve the traditionally dire prognosis for mesothelioma patients is to catch ...inhibitor) prevents the growth of new blood vessels required to sustain growing tumors. Studies have shown ...

  17. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...Addgene Executive director, Joanne Kamens, recently participated in a Reddit AMA (short for “Ask Me Anything..., work-life negotiation, and mentorship. We can direct people to the AMA in the future and link to it ...an organizer/promoter was that I didn’t have the direct link to send people until the day of the AMA. This...

  18. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...all the steps and technical considerations are directly applicable to the rat [6]. Technical differences...Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins." Nature...Central PMCID: PMC3082132. 9.Del Bene, Filippo, and Claire Wyart. "Optogenetics: a new enlightenment age for...

  19. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ... developed using yeast display technology, are directed to integrins that bind the Arginine-Glycine-Aspartate...2022). By contrast, LVDP-binding integrin α4β1 required and conferred less force, leading cells to spread...integrins, αVβ3 and αVβ5, participated but were not required for cell adhesion. The results surprised the investigators...

  20. Tips for Getting a Faculty Position

    Type
    Blog Post
    ...really are selling yourself. A sales pitch that requires reading two pages or even a paragraph to get to... Simply stated, what are your aims? You could directly state your official aims, but I suggest distilling... and Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia...

  21. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...which is ideal for observing and manipulating the entire developmental cycle. A majority of studies involving...most significant advances is the sequencing of the entire axolotl genome, revealing an abundance of genes...year 2000, becoming the first plant to have its entire genome sequenced (Koornneff and Meinke, 2010). ...

  22. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ... the activity of the original, generated using directed evolution (Tei et al., 2023).  PLD can hydrolyze... doesn’t matter. Plus, for folks interested in directed-evolution methods, the activity-based selection... J. C., & Baskin, J. M. (2023). Activity-based directed evolution of a membrane editor in mammalian cells...

  23. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ...speaking early and often. Two of my jobs have indirectly resulted from someone seeing me speak in a non-interview... research, “aha” moments, and future research directions. Don’t worry if your work hasn’t been commercialized...does follow the general structure and flow of the entire session. I myself have a terrible habit of saying...

  24. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...Brainbow-1.0 in that transient Cre recombination is required to prevent constant inversion of the construct...derivatives of the fluorescent proteins, which are directly trafficked to cell membranes. Membrane trafficking... Fouquet S, Dupin E, Benosman R, Chédotal A, Beaurepaire E, Morin X, Livet J. Neuron. 2014 Feb 5;81(3)...

  25. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...guide and the transposon RNA homology both help direct the fusion construct to the new genomic locus. ...indel formation than methods relying on homology-directed repair (HDR), and insertion is scarless, unlike...where those interactions occur in the genome. This requires both targeting a specific genome location and ...

  26. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ... in a DNA double strand break (DSB), the event required for frameshift and knock-in mutations. If one ...will generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation...sequencing The only way to quantify off-target effects entirely is to perform whole genome sequencing. While this...

  27. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...advantage to using direct epigenetic modifiers? As with any experiment, your desired outcome will determine... DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of ...

  28. Important Considerations When Using AAVs

    Type
    Blog Post
    ... Is Bitter, but Its Fruit Is Sweet Time If you require a quick onset of expression, AAV may not be the...AAVs for DREADD delivery indicates ~7-21 days are required for the onset of gene expression. However, it ... As single stranded DNA viruses, AAVs normally require second strand synthesis before gene expression ...

  29. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...variable region of an antibody does not recognize the entirety of the target protein: instead, it recognizes ...between the IgG antibody and scFv. Because the entire antibody molecule is not necessary for antigen ...other markers can be injected into an animal, to directly label circulating cells or other cells that are...

  30. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...antigen of interest in the given protocol. This requires use of one of the five pillars of antibody validation...intended application — here, flow cytometry — is the desired target, using one or more of the approaches outlined...relative protein expression. Its reliability will be entirely dependent on the selectivity of the antibody used...
Showing: 541 - 570 of 910 results