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We narrowed to 798 results for: Cre

Showing: 241 - 260 of 798 results

  1. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...CIB1 to regulate botulinum neurotoxin were also created. The depositors demonstrated the utility of light-mediated...these gas vesicle proteins scatter sound waves and create ultrasound contrast. At a very low volumetric density...s lab at MIT pushed the field further with the creation of the largest miRNA sensor collection reported...al., Nat Commun, 2019. PubMed PMID: 29934631. Screening immune cell lineages in vivo using the CRISPR-...tools have been used in immunology for in vitro screens of immune cell activity. However, a CRISPR-based...gene. CHIME can also be used for high throughput screens using pooled libraries. Find the CHIME plasmids...

  2. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    ...targeted via RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly...non-specific nucleotides into the dsRNA leading to increased RNAi efficiency. Find the T444T plasmid here... used yeast display-based directed evolution to create APEX2. They have now applied the same directed ...the biotin ligase from which BioID was derived, creating a pair of enzymes that overcome both the speed...BioID, which only functions optimally at 37℃. The increased efficiency of TurboID and miniTurbo makes PL a...include protein stability, small molecule binding screens, peptide arrays, and phage display selections. ...

  3. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ... technology has been used to optimize protein secretion and expression in the biotech workhorse yeast ...Engineering disease resistance through gene stacking, increasing production of a protein of interest, and improving...genetic circuits with multiplexed guides, mini screening libraries, direct edit optimization, and more....can be invaluable for more complex editing and screening experiments in organisms that don’t yet have these...modify it as necessary. And remember, if you end up creating a really cool MoClo kit, we would love for you...Modular Toolkit for Generating Pichia Pastoris Secretion Libraries.” ACS Synthetic Biology, vol. 6, no.... of the GoldenBraid Modular Cloning System and Creation of a Toolkit for the Expression of Heterologous...

  4. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    Published
    July 20, 2021, 1:15 p.m.
    ...assaying a secreted protein, first block the Golgi apparatus with Brefeldin A to inhibit secretion and then...through the cell membrane. Formaldehyde fixation creates cross-links between lysine residues thus preserving...flow cytometry data users will typically start by creating a dot plot of FSC versus SSC that allows them ...initially gating the lymphocytes, one could then create a dot plot of CD3 (T cell marker) versus CD19 (...cells, might then gate the B cell population and create a histogram of CD19-containing cells versus total...cells in that sample. Alternatively, they could create additional dot plots of various CD markers to further...

  5. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    Published
    April 20, 2023, 4 p.m.
    ...phobias–it’s a key part of being a scientist and an incredibly useful transferable skill. Being able to stand...even in my head. What came out, though, was an incredibly calm: “Excuse me, I seem to have lost my place...any silence, to talk. That moment taught me an incredibly valuable lesson: Your presentation will be better...your slide deck gets uploaded. Your mic emits screeching feedback. You’re muted on Zoom. Your video conferencing...provide quick updates during a company meeting increases visibility of your work, opens up collaboration...Conflict Resolution Transferable Skills Guide: Creativity...

  6. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...may not result in any cells, or all the colonies screened might not contain the correct plasmid. There’s...and if the sequence has high GC or AT content, increase the overlapping region of the primer to 40-60 ...buffers used in the previous reactions. This will increase your chances to have a successful and ultimately...pick the big colony and you are safe! Colony screening Pick a few colonies (5-10) and grow them in a ... you've identified potential clones after the screening, sequence the plasmid. Many vectors contain sequences...can use the chromatogram to spot whether any discrepancies between your sequencing result and the expected...

  7. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...2024). It reduces the amount of time to design and create a new vector to four days for an experienced bench...takes a lot of the design work out of the plasmid creation process. Each module of Fragmid represents a class...caption from McGee et al., 2024, used under the Creative Commons CC BY-NC-ND license.   Once you have... system by sequencing 60 individual assemblies created over five months. When they tested those clones... plasmids were perfectly correct. Because they created the assemblies in duplicate, they were able to ...Doench lab share this new tool and hope it helps you create great plasmids for your next CRISPR experiment....

  8. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...further modified to create ultrapotent PSEMS (uPSEMs) that have drastically increased potency. Download ...signaling pathways and can lead to such responses as decreased heart rate (Redfern et al., 1999) or altered neuron...firing. Other DREADDs, such as the KORD receptor, increase this threshold and silence neurons. DREADD-activated...actuator modules, were developed with the goal of creating receptors with less endogenous activation and ... Higher potency means lower drug doses, which decreases the risk of off target effects. These synthetic...

  9. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...consider scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size. Opting ...viral genome into additional plasmids thereby increasing the number of recombination events that are required...some cases, titers can drop 10-fold for every 2kb increase in insert size. To overcome this, lentivirus can... lentivirus routinely, you may opt for simply increasing the number of 10 cm2 dishes. While this option...handling multiple dishes will take longer and may increase the risk of mistakes or contamination. Finally...

  10. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...plasmid contains all of the desired gRNAs. This increases the chance that all the edits you want to make... cleave outside of their recognition sequence, creating flanking overhangs. These overhangs can be customized...Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing wt humanized Cas9...the gRNAs. gRNAs are flanked by glycine tRNAs to create polycistronic glycine tRNA-gRNA (PTG) constructs...using 3 distinct spacer-repeats, you could easily create 7 unique arrays (e.g. for spacers A, B, and C, ...is possible with efficiency >70%. CRMAGE is an incredibly fast protocol, with only 5 hours incubation time... Find plasmids from this paper at Addgene. 11. Cress, Brady F., et al. “CRISPathBrick: Modular Combinatorial...

  11. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...describes going through two rounds of phage display to increase the enrichment of target-specific sybodies. Final...The enriched sybody pools from the phage display screens are subcloned (using FX cloning) into the expression...

  12. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...Tsuboi et al.), limiting its use. Nonetheless, its creators noticed a few properties that have now become ... be. Cytotoxicity, improper localization, and decreased functionality are all possible consequences of...al.) Monitor gene expression in the developing pancreas by placing FP timer DsRed-E5 under the control...control of Neurog3 (a gene controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish...you planning on using an FP timer in a new and creative way? Let us know in the comments section below...differentiates human IPSCs into glucose-sensitive insulin-secreting beta-like cells." Nature communications 7 (2016...

  13. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ... second FP, often decreasing the fluorescent signal from the first FP and increasing the signal from the...an FP to a transmembrane protein significantly increases the effective concentration of the FP. The FP ...to a plane and generally only rotates in 360°, increasing the probability that two FPs will collide with...polymer formation as fusion protein concentration increases. The polymerized fusion proteins that form as ... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum and live...

  14. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    Published
    July 1, 2022, 1:15 p.m.
    ...well for everyone else?  In truth, people are incredibly varied in how they prefer to consume information...matter will drive the kinds of communications you create. Wildlife biology and ecology can make for beautifully...Once you’ve answered the above questions, you can create your short list of options and then pick whichever...Medium to high Uses visuals, audios, and text. Incredibly immersive and flexibility to visualize and contextualize...format. Equipment, expense and expertise needed can increase exponentially with complexity. Podcasts Medium...audience and communicators. Well-developed content creation platforms.  Algorithms can make it difficult ...

  15. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...Figure 1: Overview of Sanger sequencing technology. Created with BioRender.com. Sanger sequencing was considered...original Sanger procedure to improve throughput by increasing the number of reactions. The main steps of short-read... Overview of short-read sequencing technology. Created with BioRender.com. After sequencing results...3: Overview of Nanopore sequencing technology. Created with BioRender.com. Nanopore can provide longer...time, but does not leave much flexibility for discrepancies in backbone sequences. Most theoretical sequences...identity and length, but disregards the specified discrepancies. Any sequences that are a perfect match from...

  16. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    Published
    Nov. 3, 2016, 2:30 p.m.
    ...humanities, and 32% in the social sciences go uncited. Creative titles and controversial topics can garner some...Article, since the 1980s researchers have been increasingly citing research from outside of their disciplines...interdisciplinary research while at the same time increasing interdisciplinary citations in their own work...instant sharing across multiple other sites), I created a new platform for sharing video abstracts about...effectively.” How to share your science through video Creating an effective video about your research is not ...offers a 5-step process and several examples for creating effective research videos.  Karen McKee, also...

  17. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ... I’ll describe an episomal vector system that increases reprogramming efficiency through the expression...requires stringent biosafety measures, thereby increasing the costs of the procedure. Episomal reprogramming...reprogramming is an ideal and cost-effective method for creating clinical-grade, safe, nonviral and nonintegrating...episomal reprogramming cocktail by Okita et al. increased the reprogramming efficiency in PBMCs [7]. The...transiently. The transiently expressed EBNA-1 increases protein expression from the other episomal plasmids...used on PBMCs, the pCXLE episomal vector system increases the efficiency of reprogramming from 0.001% to...

  18. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...inhibitor (UGI) minimizes the second outcome and increases the generation of the desired T-A base pair. ...act on DNA for adenine base editing. They first created a defective chloramphenicol resistance gene (CamR...antibiotic resistance. To find such a protein, they created a mutant library of E. coli tRNA adenosine deaminases...into E. coli harboring the defective CamR gene. Screening for surviving colonies and subsequent rounds of...genotyping. If successful base editing causes the creation or destruction of any restriction enzyme (RE) ...

  19. 2021 Deck the Lab Winners!

    Type
    Blog Post
    Published
    Jan. 25, 2022, 2:15 p.m.
    ...Saves Christmas! created by the Moore lab at the Van Andel Institute. Krampus was created by Nate Levine...Warrior The Moore lab, who was full of holiday creativity, also receives an honorable mention for their...2021   2nd Place: The Winter and the Falcon Tube created by the team efforts of the Yit Heng Chooi Lab at... December 4, 2021   3rd Place: O! Beaker Tree Created by Joanne Jary from Lab Support at the John Innes...

  20. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ...DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve as both a ...considerations for designing a repair template to create mutations HDR donors can contain insertable sequences...circular plasmid templates is generally low; to increase the frequency of edits, researchers have designed...complementary ssDNA. Easi-CRISPR was shown to increasing editing efficiency from 1-10% with dsDNA to 25...Frequency of Homology-directed Repair (HDR) for Increased Efficiency of CRISPR/Cas-mediated Transgenesis...
Showing: 241 - 260 of 798 results