We narrowed to 798 results for: REP

... bioinformatic screens that looked for a CRISPR repeat array near a putative effector nuclease. The Cas13d...localization signal (NLS). In a HEK293 mCherry reporter assay, CasRx and AdmCas13d produced 92% and 87%...processed gRNA array (22 nt spacer with 30 nt direct repeat) mediating potent knockdown. Multiplexing from ...Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional Activators...

...immunotherapy, as well as new approaches to reprogramming tumor-associated inflammation for therapeutic...reflect how cancer/immunology researchers are preparing and carrying out this multi-pronged attack. Inflammation...This immunosuppressive environment in turn may represent a favorable state for tumor progression (Ref 2...potential to become an open-source pipeline and data repository for cancer researchers. Go open-source! Impressions...

... et al used four oligos that consist of direct repeats and crRNA.  Similar to a jigsaw puzzle, the oligos... reverse the orientation of the crRNA’s direct repeats when expressing the Cpf1 multiplex system from ...virus.  Reversing the orientation of the direct repeat protects the (+) stranded lentivirus RNA from Cpf1...This undoes the original reversing of the direct repeats, leading to expression of a crRNA array that can...

...accurate, IPTG binds and inactivates the lac operon repressor, thereby allowing lac expression). When expressed...multiple cloning site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left)...-complementation (cell B). As shown on the representative plate to the right, colonies with an insert-containing...defined medium, without which the cells cannot reproduce to form colonies (used to complement auxotrophic...

... get started with mentoring. Read this Catalyst Report “Making Mentoring Work” to learn about what makes...mentoring can work if both parties are diligent about replying and thinking about good questions and answers....Thank them anyway and move on. If they say yes, be prepared with questions and topics to discuss. The mentee...make your mentor want to keep working with you is report back with progress. Tell them how their advice ...

...break in the genome anywhere you would like. Cells repair those breaks in one of two ways: by putting the... a gene. One can also exploit homology-directed repair by flooding the cell with a mutant template and...and fooling the cell into using that template for repair. By using these two scenarios, you can either knock...where particular variants or mutations have been reported in people, we can use this kit to knock in those...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo toolkit for... standardized parts, it allows for greater reproducibility as scientists can assemble parts from different...sequences for genomic integration by homology directed repair. Using parts from this kit, the McCormick lab generated...

...little was one minute BASU generated a strong streptavidin signal (~250-fold enrichment) when compared ...Ramanathan et al. also used the Contamination Repository for Affinity Purification (CRAPome), to identify...identify highly biotinylated background proteins reported by other research groups that performed BirA*-based... that QKI is a host protein critical to Zika replication. Human genetic disease. RaPID can also be used...

...conditions, i.e. with low excitation power. By repeating these measurements for different fluorescent proteins... be noted that the photostability measurements reported in the literature are performed in different ways...information can help guide your understanding of reported data. Photobleaching results in a loss of fluorescence...a set-up has been changed (for instance after replacing a broken lightbulb or exchanging excitation filters...

...How do microenvironment enzymes and metabolic reprograming influence tumors development? Being pathological...cells improve tumor’s durability, while others repress it. Unfortunately, as a whole, microenvironments...microenvironments promote tumor durability instead of repressing them, making it even harder to cure cancer. To understand...affect tumor progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential for a...

...AATTGXXXXXXXXGAATAGGTCTTCTCGAAGACCCAAACNNNNNNNNNNNNNNNNNNNC -3’, where NNN and XXX represent the 20-bp gRNA target sequence and its 8-bp barcode...and applications in functional genomics, cell reprogramming, and beyond. For further details, please check..."Genome-scale CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. ... for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683. PubMed PMID: 26190106. PubMed...

...sigh. The problem is that many of these studies report correlations without clearly testing whether a ... when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites blocked by dCas9....for dCas9 to interfere with DNMT1 even through replication. The dCas9 also needs to be removeable, otherwise...Targeted DNA demethylation in vivo using dCas9–peptide repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol...

...for genome editing, activating gene expression, repressing gene expression, and much more. But there’s one...meningitidis (Nme) (Pawluk et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and... viruses. For example, an Acr gene was used to replace genes in the difficult-to-engineer Sulfolobus islandicus...viral particles that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-...

...mild symptoms. This changed in January with the report of an outbreak of the virus in Brazil that was ...-ready format when the Zika virus outbreak was reported, and we felt that, given the critical need for...such a short timescale. Our article ultimately reported the development of a rapid design and prototyping...Find the Zika Sensor Constructs at Addgene As we report, we have embedded this next generation of toehold...

...region. Thankfully, due to their prominence, the preparation of the fiber optics is probably the least difficult...effect is also an important consideration when preparing the implant itself, which brings me to the second...same considerations apply to the polishing and preparation of the patch cable (although you may be able ...wirelessly-controlled optogenetic implants have been reported by several groups [4, 5, 6]. These have the obvious...

...assertive communication.” - Jim Rohn, Author, Entrepreneur, Motivational Speaker Do you have a difficult...supervisor might expect weekly or biweekly progress reports. However, a hands-off busy supervisor might get...update, and whether he or she would like a written report or to meet in person.   Regardless of your supervisor...for help so you can get the support you need. 2) Prepare an agenda for every meeting Supervisors are busy...

...order to head to Ireland and take a chance as entrepreneurs. To make things more interesting, we are working...career, but it was enough to start us down the entrepreneurial path. We threw some of our own money at this...genetically modified organisms, and GMOs have a bad reputation. The public is can be fearful of this technology...thinking about making the switch from academia to entrepreneurship, share your thoughts with us in the comments...

...transfection). This differs from typical insect cell preparations, which we talk about below. Processing The viral...eliminate any HEK-293T cells. If you titrate your viral preps, lentiviral titration requires an additional couple...to collect your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in...Lentivirus Production Tips for Titering Your Lentivirus Preps More resources on Addgene.org Addgene's virus protocols...

...annealing and extension is repeated 25-35 times to exponentially replicate the target DNA of interest....concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify DNA sequences ...Quantify RNA levels Molecular Diagnostics NGS library prep QC ddPCR Determine ultrasensitive and absolute...

...Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's...FP dimerization. Equally problematic, several reportedly monomeric FPs are not actually monomeric, at ...Incorporation into a FRET biosensor A FRET biosensor is a reporter that undergoes a conformational change to bring...help you make better fluorescent fusion protein reporters for your studies and greatly lower your chances...