We narrowed to 381 results for: BRE
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TypeBlog Post...endonuclease Cas9 to create a DNA double stranded break (DSB) at a genomic site specified by a guide RNA... sequence. Users can design gRNAs to induce the break at a specific genomic site and, using the endogenous...homology directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the cell ...
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A Guide to Starting Your Own Journal Club
TypeBlog Post...multiple teams or to stay abreast of current trends in the literature, sampling a breadth of work from a field...subsequent meetings. The number of participants, breadth of the discussed topic, and level of general busy-ness... -
Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design
TypeBlog Post...to account for the effects of Cas9-induced dsDNA breaks. Are off-target cutting patterns in genome-wide...control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting vs. safe-targeting...inherent effects, or background noise, of dsDNA breaks caused by CRISPR. See supplemental figure 6 for... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog Post...CRISPR/Cas9 edits that rely on double strand DNA breaks. Due to this inefficiency it is crucial for scientists.../10.1093/nar/gkaa124 Martin AS, Salamango DJ, Serebrenik AA, Shaban NM, Brown WL, Harris RS (2019) A panel...s41598-018-36739-9 St. Martin A, Salamango D, Serebrenik A, Shaban N, Brown WL, Donati F, Munagala U, ... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog Post...single-stranded nick. In order to produce a double stranded break, two sgRNAs must be used to target two Cas9-nickases...achieve your precise edit without even making a break. Base editors are dead Cas9 (dCas9) or nCas9 enzymes...converts a C to U without producing a double strand break. Prime editors are the second generation of... -
Transferable Skills Guide: Creativity
TypeBlog Post...crunch, your break can be as simple as a few minutes of meditation or a walk to the break room for a sip...going throughout these challenging times. Take a Break! If all else fails, one of the best things you can... -
Mesothelioma - Causes, Symptoms, and Treatment
TypeBlog Post...the abdomen. Long story short, when asbestos is breathed in (or less frequently, ingested), it can get ...like lung (about 224,000 diagnosed this year) or breast cancer (about 246,000 diagnosed this year). Research... to detect mesothelioma earlier than ever. From breath tests to blood tests, researchers are hopeful that... -
Five Popular Model Organisms, Part 2
TypeBlog Post...the rat (Gibbs, et al., 2004). Additionally, breakthroughs in gene modification tools and methods such ...organism; they are relatively simple to maintain and breed and they have large, easily modifiable embryos, ...frog research is The National Xenopus Resource, a breeding facility for X. laevis and X. tropicalis, the ... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog Post...select for desirable traits using traditional plant breeding approaches, these techniques are cumbersome, often...repair, these methods do not involve double strand break (DSB) formation and do not require donor DNA, . ...Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog Post...you obtained the desired chimeric mice, you would breed them many times, hoping to produce a mouse with ...need to remove the selection marker, likely by breeding to a mouse line with a recombinase. If it turned... insert two loxP sites. Since two double-strand breaks are made, it is common to see deletions of the ... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog Post... is usually accomplished by Cas9-mediated dsDNA breaks: following a cut, the error-prone nature of non-homologous...although this is rarely done). When targeting a dsDNA break for HDR, the choice of target site is far more constrained...which makes DNA changes in the absence of dsDNA breaks (Rees et al., 2018). For C>T and A>G base editors... -
Antibodies 101: Multiplex Immunofluorescence
TypeBlog Post...marker like the Hoechst stain for the nuclei. (See Brewer et al, 2022 for a sample protocol.) Simultaneous... Nguyen, K., Van Der List, D., Murray, K. D., Engebrecht, J., & Trimmer, J. S. (2019). A toolbox of IgG... 8, e43322. https://doi.org/10.7554/eLife.43322 Brewer, M., McDonough, L., Zhu, Y., Neumann, E., De Caestecker... -
An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog Post...homology arms of 35-80 bp, and DNA double-strand breaks up to 54 bp from the desired insertion site result...knock-in efficiency, presumably by channeling DNA breaks into the homologous recombination repair pathway...interesting to note that recipients for the 2015 Breakthrough Prize included Jennifer Doudna and Emmanuelle... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog Post...development (Grimm & Lavis, 2022). With the wide breadth of fluorescent tools that have been developed throughout...is an important consideration for assay design (Breen et al., 2016). When selecting dyes, it's important...21842. https://doi.org/10.1038/s41598-020-78827-9 Breen, C. J., Raverdeau, M., & Voorheis, H. P. (2016).... -
Antibodies 101: Flow Cytometry
TypeBlog Post...protein, first block the Golgi apparatus with Brefeldin A to inhibit secretion and then follow the typical...NK) cells. Tips for flow cytometry success Break up cell clumps Success in a flow cytometry experiment...the sample through a nylon mesh cell strainer to break up clumps before running on the instrument. To sustain... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...al. 2022. Image created with BioRender.com. Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory.../ENEURO.0373-21.2022 Brown, D., Altermatt, M., Dobreva, T., Chen, S., Wang, A., Thomson, M., & Gradinaru...Kumar, S. R., Miles, T. F., Chen, X., Brown, D., Dobreva, T., Huang, Q., Ding, X., Luo, Y., Einarsson, P... Kumar, S., Miles, T. F., Chen, X., Brown, D., Dobreva, T., Huang, Q., Ding, X., Luo, Y., Einarsson, P... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...other organisms, CRISPR-induced double stranded breaks are lethal because the non-homologous end-joining...ineffective in bacteria, CRISPR-induced double stranded breaks (DSB) are lethal. Addgene depositor Luciano Marraffini...genomic locations, and introduces double strand breaks, the bacteria are unable to resolve them, and the... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog Post...HDR), the pathway for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared to non-homologous...Base editors chemically modify DNA bases without breaking the DNA backbone, sidestepping the problem of ...converts a C to U without producing a double strand break. UGI = uracil DNA glycosylase. Created with BioRender.com... -
Addgene’s New Way to Help You Share Science
TypeBlog Post...talk to scientists from around the world making breakthrough discoveries!” I knew right then I had found .... Summaries of the latest technologies and breakthrough research. Insights about how to navigate through... -
Podcast: A Malawian Professor's Path to Biotech Research
TypeBlog Post...Horticulutre Biotechnology, he did his PhD in Plant Breeding, Genetics, and Biotechnology at Michigan State...collaboration across the globe. Listen here! Podcast Breakdown (Time: Topic) 0:00-1:00 : Intro 1:01-3:48 : Kingdom...