We narrowed to 777 results for: REP
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TypeBlog PostPublishedJan. 26, 2016, 3:30 p.m....often used for animal germline modification, to repair or add in disease-causing mutations, but, until...suited for genome editing. The homology-directed repair (HDR) pathway is downregulated in mature tissues...Although the three DMD studies referenced here represent a great step forward for CRISPR gene therapy, ... and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit Genes Read How Non-Homologous End...
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Antibodies 101: The Basics of Western Blotting
TypeBlog PostPublishedMarch 8, 2021, 2:41 p.m....addition, PVDF membranes can be repeatedly stripped of an antibody and reprobed with different antibodies allowing... (SDS-PAGE) is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content...control. In these cases, the western should be repeated. Finally, while westerns are a useful way to ... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog PostPublishedSept. 24, 2020, 1:15 p.m....mutation, one generally relies on homology directed repair (HDR) to incorporate new information into DNA. ...cut site is >30nt from the proximal ends of the repair template (Yang et al., 2013). This means that, ...the authors note that “clonal heterogeneity may represent a more serious obstacle to the generation of truly...efficiency of CRISPR/dCas9-mediated transcriptional repression. Nucleic Acids Res 44:e141–e141 . https://doi.org... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog PostPublishedApril 16, 2024, 1:15 p.m....Live-cell labeling strategies. The green structure represents a fluorophore or other useful ligand, gray structure... self-labeling tools: Tetracysteine: The first reported chemical labeling tag, from Roger Tsien’s lab ...self-labeling fusion tag based on the human DNA repair protein AGT that gets covalently linked to derivatives...collection. CLIP-tag: Also in 2008, Kai Johnsson’s lab reported another AGT-based tag named CLIP-tag, which reacts... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog PostPublishedOct. 17, 2019, 12:28 p.m....infect defined neuronal subpopulations, and this represents a major downside in optogenetics (Belzung et ...gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to easily detect opsin-expressing...behaviors (de Jong et al., 2019). FLEx vectors represent, therefore, an ideal partner to optogenetics to...recombinase in mouse primary afferents." Scientific reports 8.1 (2018): 7321. PubMed PMID: 29743652. PubMed... -
Plasmids 101: Modular Cloning Applications and Kits
TypeBlog PostPublishedMay 14, 2024, 1:15 p.m....sub-cellular localization signals, plant markers, and reporters (Engler et al., 2014). This kit is a great starter...into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing of sgRNAs...generic MoClo system for any given species can be repurposed for CRISPR applications (gene insert = Cas9 +...Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog PostPublishedMarch 21, 2024, 1:15 p.m....ligand-binding assay. Scientific Reports, 6. https://doi.org/10.1038/srep25769 Grimm, J. B., & Lavis, L. D....ligands for dopamine D2/D3 receptors. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-78827...mitochondrial dynamics in mammalian cells. Biophysics Reports, 3(4–6), 64–72. https://doi.org/10.1007/s41048-... -
Typing CRISPR Systems
TypeBlog PostPublishedMarch 18, 2025, 1:15 p.m....subtypes, A–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first...adaptations for increased fidelity, activation/repression, high precision edits, and more. Figure...Schematic of the basic CRISPR-Cas9 mechanism, representing type II-A. Created with BioRender.com. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named A–I and... -
CRISPR Cheat Sheet
TypeBlog PostPublishedJan. 27, 2021, 2 p.m....Clustered Regularly Interspaced Short Palindromic Repeats: Sequences of DNA found in bacteria that allow ...CRISPR can be used to cut the DNA sequence. With a repair template, CRISPR can be used to introduce a functional...deaminase can convert adenosine to inosine. Activate/Repress: Modified versions of CRISPR nucleases that can... -
2018 #DeckTheLab Contest Winners
TypeBlog PostPublishedJan. 15, 2019, 2:19 p.m....this time of year 😁 we stopped working on our reports/lab stuff for a few minutes to create this lovely...Matthew J. Sikora (@mjsikora) December 18, 2018 Fireplace from the Dyer Lab If you’re feeling as chilly ..., they turned one of the lab computers into a fireplace scene. Got our @EmoryChem lab office looking... -
Visualizing Your Plasmid Request Data
TypeBlog PostPublishedApril 11, 2023, 4 p.m....grant proposals or to demonstrate impact in annual reports. While the raw data is, of course, interesting ...data, including total requests, total countries represented by requestors, year of first reagent sent, year...and the source code is available in the Github repository (you can find the link in the upper right hand... -
The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing
TypeBlog PostPublishedMarch 10, 2015, 2:25 p.m.... Bank (DSHB), directed by David R. Soll, is a repository for monoclonal antibodies, distributing over ...against new cancer biomarkers. We believe that repositories, such as Addgene and the DSHB, play an important... to ensuring affordable access to materials, repositories provide standardized quality-control and technical... -
500,000 Plasmids Shipped and Counting!
TypeBlog PostPublishedMarch 26, 2015, 3:24 p.m....more scientists outside of the US now use the repository – about 50% of plasmids are shipped to labs outside...manage our website, and continue to improve the repository. We love being able to provide a useful (and ...being part of our growing community! Our plasmid repository succeeds because of our amazing community. The... -
Addgene's Website Update
TypeBlog PostPublishedApril 8, 2024, 2:15 p.m....two options: “Search Repository” and “Deposit Materials”. The “Search Repository” function is our same...Antibody of the Day! Figure 4: The plasmid, viral prep, and antibody of the day While those are the biggest... -
Your Top Requested Plasmid in 2015!
TypeBlog PostPublishedDec. 30, 2015, 3:30 p.m....We’ve dug into the data from our repository to find our most requested plasmid in 2015. In line with ...that the plasmid most requested from the Addgene repository in 2015 was... lentiCRISPR V2 LentiCRISPR...fantastic research tool has been requested from the repository nearly 1,000 times in 2015! If you haven’t already... -
A Conference By Postdocs For Postdocs: Future of Research
TypeBlog PostPublishedSept. 2, 2014, 3:05 p.m....published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates. Scientists must do science ...that too few of these trainees are sufficiently prepared during their 6-12+ (!) years of training to get... finding ways to provide new types of career preparedness training for their scientists. I have worked... -
Your Top Requested Plasmid in 2016!
TypeBlog PostPublishedJan. 11, 2017, 8:47 p.m....types. Purpose SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from a Cbh promoter. Cbh ...un-tagged versions of SpCas9 and further variants replace SpCas9 with SpCas9n (nickase), which nicks DNA ...breaks and may better facilitate homology directed repair. You can find all of these variants, as well as... -
Designer PUF Proteins for Any RNA Target
TypeBlog PostPublishedJune 17, 2014, 7:57 p.m....Urbana-Champaign. The PUF proteins include eight modular repeats, each of which binds a consecutive base in its ...colleagues led by Zhao described and demonstrated in a report published in The Journal of Biological Engineering...Engineering in March is fused to a translational repressor. In principle, however, designer PUF proteins can... -
Our Most Popular Blog Posts from 2019
TypeBlog PostPublishedDec. 31, 2019, 2:26 p.m....plasmids” from last March were quite popular and represent the diversity of tools we share on behalf of scientists...and guest blogger Erik Snapp breaks down how to prepare a chalk talk for an academic job interview including... blog posts Resources at Addgene Browse the repository Find educational resources at Addgene Search ... -
What's New in CRISPR - December 2019
TypeBlog PostPublishedDec. 17, 2019, 2:37 p.m....deaminase (TadA) to a catalytically impaired Streptococcus pyogenes Cas9. They called this editor Target-ACE...ribonucleoproteins for transient expression. They replaced the tetraloop of the sgRNA scaffold with a com...adenosine-to-inosine (A-to-I) RNA editor called REPAIR by fusing catalytically inactivated RNA-targeting...