We narrowed to 965 results for: REP

...nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an activation domain (CRISPRa) to...CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While CRISPRko... protein (dCas9). dCas9 is then tethered to a repressive domain, such as KRAB, to prevent efficient transcription...Dolcetto library! Now also available as a lentiviral prep. Calabrese: human CRISPRa sgRNA library In addition...Find the Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench CRISPR..."Genome-scale CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. ...

... in cancer studies, act as a reporter for DNA double strand break repair, and to label pathogenic intracellular...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...been done to expand the red-fluorescent protein repertoire; however, these proteins are unique from GFP ...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...

...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. Today... set of fusogenic and targeting components to reprogram delivery vehicles like lentiviral vectors, CreVLPs...doi: 10.1038/s41592-023-01880-5. PMID: 37188954. Preprint available on bioRxiv – doi: 10.1101/2023.03.09.531939...doi: 10.1016/j.cell.2023.05.044. PMID: 37385249. Preprint available on bioRxiv – doi: 10.1101/2023.02.07.527548...doi: 10.1038/s41587-022-01524-7. PMID: 36593408. Preprint on bioRxiv — doi: 10.1101/2021.10.13.463862.  ...

...in our repository that you can use to study these genes! Groucho, a transcriptional co-repressor The first...after the protein Groucho, a transcriptional co-repressor important in development. Groucho family proteins...specific set of DNA-binding transcription factors to repress the expression of genes primarily involved in development... see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman and... Groucho/TLE/Grg family of transcriptional co-repressors." Genome biology 9.1 (2008): 205. PubMed PMID...

... strand break (DSB) mediated homology dependent repair by enhanced green fluorescent protein (EGFP) reconstitution...sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination; SSA, single strand...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using...Samantha Young is a Medical Writer with a PhD in reproductive biology and genetics. She loves to communicate...expressing Cas9 and single guided RNA." Scientific reports 3 (2013). PubMed PMID: 24284873. PubMed Central...Non Homologous End Joining and Homology Directed Repair Additional Resources on Addgene.org Find Software...

...Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue...goes, “failing to plan is planning to fail.” Preparation is a key step to any experiment and can help ...experimental details on the spreadsheet: Name your viral prep Enter the specific names of the plasmids you plan...using 28 μg of total plasmid (Transfer, helper, and RepCap added together) per 15 cm dish or 2500 μg of plasmids...

...dCas9-VP64) or repressors (dCas9-KRAB or dCas9-SRDX) can also be used to activate or repress target genes...engineering CRISPR can be used to knockout, activate or repress target genes in plants using the same general experimental...containing plasmids for knockout, activation and repression of target genes in plants and many of the aforementioned...in human cells. In contrast to homology directed repair, these methods do not involve double strand break... the Patron Lab.  As with all plasmids in the repository, we highly recommend reading the associated publications...

...regulatory elements (to mutate promoters/enhancers in reporter constructs), in functional analysis of proteins...introduced to endogeneous DNA through homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded ...to the endogenous target, yet to facilitate the repair, the template should be resistant to Cas9 cleavage...your primers should be free of palindromic and repetitive sequences, but if present, a minor extension ...product (see figure). Template A high purity plasmid prep significantly increases the success rate of site...’ll want to steer clear of these strains when preparing the plasmid template for site directed mutagenesis...

...similar protein that was discovered in the 1960s, strepavidin. The Biotin-avidin complex can then be used and...primary antibody respectively. Avidin bound to a reporter is then added to the reaction for easy detection... experiments. During detection, a biotinylated reporter enzyme that is pre-incubated with free avidin ...protein of interest leading to amplification of the reporter signal and easier detection. This strategy is ...Forster, A. C., Skingle, D. C., & Symons, R. H. “Preparation and uses of photobiotin.” Methods in Enzymology...mammalian cells with biotin ligase and monovalent streptavidin.” Nature Protocols 3.3 (2008): 534–545. PubMed...

...versions obsolete, the techniques we use in lab are replaced by improved versions that save both time and money...observed. Today Southern blotting has been largely replaced by real-time PCR to answer the same experimental...researcher (or a simple computer program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility...this technology, radiolabelled nucleotides were replaced with fluorescently labeled ones allowing for fluorometric-based...discoveries (5, 8). While newer techniques have largely replaced these, these techniques still have useful applications...already commercially available although they are reported to have high error rates (10). It is projected...

... our viral vectors repository this year (and it's only March!)  In our repository, you'll find the latest...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 21...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to view viral vector reports from other users. And don't forgot to use the link...

...research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649    ... to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles in non-elite...publications. (As an interesting aside, Nature recently reported that the most cited work in history is a 1951 ... the best way to measure impact? As that Nature report also noted, papers detailing Nobel Prize-winning...

...gradually diminish over time; and, importantly, repeated excitation generating reactive oxygen species ...difficult-to-study genes. Its main function is to regulate DNA repair, senescence, and apoptosis. Work by Kamio et al...the authors used a p53 response-element NanoLuc reporter to show that the mutants abolished transcriptional...This work highlights HiBiT tagging as a sensitive reporter for tracking proteomic cargo flow via EVs. Find...., & Wood, K. V. (2012). Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone...carcinoma in human tissue and in vitro. Oncology Reports, 50(5). https://doi.org/10.3892/or.2023.8631 Tsien...

...plasmid" section Anytime you use a plasmid from the repository, we request that you cite it using both its Addgene...find the fruits of your labor and won’t have to repeat the modification. This will save both time and ...you to browse through the many plasmids in the repository, our scientists have curated a number of special...attention to the great variety of plasmids in the repository. And last, but certainly not least, we often ...Special Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

...before using an antibody.  Antibodies and Reproducibility First, let’s review why validation is so important...called out antibodies as major culprits in the reproducibility crisis of biomedical research (Baker, 2015;...proposed 5 Pillars of antibody validation which represent five experimental strategies researchers can use...benefit and we can all help improve antibody reproducibility!     References and Resources Additional ...plasmids here!  References Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature ...10.1038/521274a  Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...of a plasmid, the first step is always to send prepped plasmid DNA for next generation sequencing. We ...promoters, selectable markers, tags, origins of replication, and fluorescent proteins that are already included...sequence by NGS. These can include GC-rich and repetitive regions. These regions require additional verification... plasmid off for Sanger sequencing.     If a repetitive region is present in the plasmid, we might need...analyze sequencing results for plasmids in the repository at a glance. Our Snapgene-powered plasmid maps...

...ADAR constructs from the REPAIR (RNA Editing for Programmable A to I Replacement) system. Like RNA targeting...Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed Repair and 3 Tips to Improve HDR Efficiency...Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair mechanisms: CRISPR 101: Non-homologous End Joining...

...behaving mice to gather the following types of data: Report time-course and concentration of released DA in...optogenetically activated and inhibited neurons in live mice Report modulation of DA signaling during learning and ...moving mice to gather the following types of data: Report endogenous DA release from a single dopaminergic...looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over the course of Pavlovian...of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

... Specifically, viruses are able to infect and replicate in host cells, but they are susceptible to cells...transposon consists of a mirrored set of inverted repeats (green) flanking the gene of interest (blue) in...is expressed (red stars) and binds the inverted repeats (green); and an endonuclease reaction occurs which...puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each contain...region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new tool kit allows...

.... In doing so, the dimer shifts from one that represses transcription to one that activates transcription.However...BLADE The team modified AraC to create BLADE by replacing its dimerization domain with VVD, a light-responsive...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...contains BLADE, the PBAD promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid... plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY:...