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We narrowed to 967 results for: Cre

Showing: 401 - 420 of 967 results

  1. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...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...

  2. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...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...

  3. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ... 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...

  5. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    ...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 ...

  6. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...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) ...

  7. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ... 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...

  10. 2021 Deck the Lab Winners!

    Type
    Blog Post
    ...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...

  11. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...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...

  12. Evolution of Lab Techniques

    Type
    Blog Post
    ...including easier automation, higher throughput screening, and a lower requirement for the amount of DNA...transgenic animals may choose Southern blotting to screen founder transgenic animals for the inserted transgene...DNA sequence content by comparing the fragments created by different restriction enzyme digests. Incubating...specific fragments of a DNA sequence of interest to create a physical map of the DNA. Starting in the 1970s...below), restriction mapping may still be used to screen multiple colonies before selecting a smaller subset...allowing for fluorometric-based detection. This increased sequencing speed ultimately leading to what are...

  13. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ... associated hybridoma cell lines. The DSHB was created by the National Institute for Child Health and ... which are each produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells ...individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing a ...and protein tags (c-MYC and HA). We have also created the Monoclonal Antibody Research Institute dedicated...partially humanized rabbit antibodies that exhibit increased avidity to high profile antigens; and 4) generating...

  14. Providing Plasmids to Researchers in Developing Countries: Addgene and Seeding Labs Team Up

    Type
    Blog Post
    ...donors to supply surplus lab equipment to carefully-screened research and teaching departments at universities.... Like Seeding Labs, Addgene also feels it’s incredibly important for scientific breakthroughs, reagents...scientific community and are always taking steps to increase plasmid distribution and disseminate knowledge... we need to expand our efforts to find new and creative ways to reach developing countries. As part of... eliminate much of white space on this map and increase plasmid sharing across the globe! We hope that...

  15. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ...use the dropdown to increase the length of the list (purple oval in the screenshot below). You can also...plasmid page. Check out the purple arrow in the screenshot below to see what I mean. Additional features...list of papers citing your deposited plasmids Increased visibility of your plasmids in the "Associated...listed in the "Articles Citing This Plasmid" list increasing your work's visibility Help us improve our citations...

  16. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    ...of protein activity. Together, these limitations create challenges for measurements needed to move discoveries...means to quickly answer research questions before creating knock-ins.  Figure 1: Schematic of HiBiT...Functional assays further showed that imported ADAR1 increased oncogenic editing of its most common target, AZIN1...UBE3C. Degradation of CFTR was measured through decreases in HiBiT signal, which was slowed by knockdown...factors present unique challenges for study. These secreted signaling proteins regulate vital cell and tissue...physiologically and endogenously relevant models, tools that increase confidence become more important. For many difficult-to-study...

  17. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide...selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating knockout models...for generating mouse models Although CRISPR is incredibly useful for generating mutations by NHEJ and generating...CRISPR, one must beware that not all gRNAs are created equal. Some work better than others. There are ...Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR...

  18. Science as Art

    Type
    Blog Post
    ...information on black screens, but not to know even the life cycle of the creature they deal with.” So ...Wildlife Magazine. It is easy to see how these incredible images are attracting so much attention. When...efforts, as the subject matter is seen to have increased value and is appreciated by a wider audience for... by holding scientific art competitions where creations can go on display for all to enjoy. The largest...

  19. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ... Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Below... paired DSBs. This method allows for deletion screening by conventional PCR. Biallelically disrupted cells...available strategies by which biologists can imagine, create, and examine novel genetic configurations. Watch...transfected cells via FACS, primer validation, and screening by PCR for biallelic deletion clones:    ... and delivery, as well as their techniques for creating deletions, identifying these deletions, and finally...

  20. What's New in CRISPR - September 2019

    Type
    Blog Post
    ... your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 activation...CRISPR knockout libraries for cancer research Creating knock-ins with GeneWeld vectors GeneWeld from ...Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins of fluorescent proteins, fusion proteins...library with SpdCas9 to perform pooled CRISPRi screens to identify essential genes and phage host factors...metastasis cohorts and a prior mouse metastasis CRISPR screen. The Chen Mouse Tumor Suppressor Gene CRISPR Knockout...
Showing: 401 - 420 of 967 results