We narrowed to 718 results for: des.1

...(RBD) of the viral spike protein.   Figure 1. (A) SARS-CoV-2 RBD DMS Library generation and sequencing...library, each plasmid encodes the mutated spike protein and a barcode, and is designed so that the SARS-CoV....1074/jbc.RA120.01289     Novel protein cage provides simple enzyme protection in yeast bioproduction...enzymes in a reaction pathway, the MPyV system provides a simple, efficient method for protecting a single...- Ampl133 targets S. cerevisiae genes with 6-12 guides per gene where gRNA expression is induced by anhydrous...

... education and scientific writing.     References 1. Martz, Colin A., et al. "Systematic identification...die from cancer-related complications each year. Despite these grim statistics, new strategies for attacking...cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations in a variety of... enabled researchers to begin discovering and designing molecules that are showing some promise in their...in the screenshot below). Currently, the page includes interactive maps for the Ras, mTor, and p53 pathways...

...for your first journal club to begin! Figure 1: Discussing next-generation sequencing at the Addgene... on single papers could be more important.  Any slides, presentations, recordings, and other materials...those who were unable to attend. We capture the desktop and audio recordings of the meeting using teleconferencing... ensure that the journal club is useful for the widest audience, you should get feedback from those who...served. I hope that with some of the information described above you feel empowered to start your own journal...

...genome engineering and microbiology.     References 1. Chen, Sean, et al. "Highly efficient mouse genome...transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains a site for inserting a custom...targeted approach using RNPs has recently been described in vitro, with the hope of eventually using this...expertise. Final thoughts The Cas9-gRNA RNP method provides scientists with an additional means to deliver...other CRISPR delivery techniques Find tips on gRNA design Resources on Addgene.org Browse all CRISPR plasmids...

...off-target effects (Zibitt et al. 2021).    Figure 1: A schematic illustration of CRISPR/Cas9-based approaches...non-coding RNAs (lncRNA). lncRNAs are over 200 nucleotides in length and lack an open reading frame, so ...transcription machinery of a target gene or DNA that codes for ncRNA. CRISPRa, on the other hand, recruits ...which targets RNA instead of DNA and selectively degrades RNA molecules. This system is often used to knock...

...which you’ve already fluorescently labeled. Figure 1: Using Split-BioID to Identify Protein Protein Interactions...specific protein-protein interactions, SplitBioID provides a candidate list of proteins that are part of ...

...article, they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several... Parts. This library delivers 2 randomly paired guides per construct and allows a reduced scale in screens...

...supervisor, even if he or she is a difficult person: 1) Understand your supervisor’s expectations Every ...familiar with the minute details of your work. Describe the problem by stating the facts and explain how...

...many mammalian vectors are designed to shuttle between species. Figure 1: Shuttle vectors are vectors...affects all kingdoms of life. Of course, that includes you. Besides the risk to the researcher, Zeocin has been...host’s replication machinery. A shuttle vector designed for unrelated bacteria should include oris for...target species. Fun fact! Plant vectors are usually designed to shuttle between E. coli and Agrobacterium tumefaciens...protects against any of them. But different aminoglycosides work on different host species: kanamycin targets...media that supplies the key nutrient. The vector provides the missing synthesis machinery, and the cells...image for individual analysis. A shuttle vector designed for this approach in a mammalian cell experiment...

...If you already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise ...components into mammalian cell lines is quite broad and includes several different methods, so we've summarized...

...and primary cells. References 1. Doench, John G., et al. "Rational design of highly active sgRNAs for CRISPR-Cas9...These libraries utilize optimized sgRNAs that were designed based on the rules identified in Doench et al....1,000 non-coding control sgRNAs. This library was designed to improve on-target activity while reducing off-target...with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello library... that if an essential gene plays a role in the desired screening phenotype, it may be missed because it's... the sgRNA metrics, confirming again that sgRNA design plays a significant role in library efficiency....CRISPRa, sgRNAs were targeted between 150-75 nucleotides upstream of the TSS. Similar to the Brunello ...

...testing. You can find Nick on LinkedIn. References 1. Skloot, Rebecca, and Bahni Turpin. The immortal life...contamination and match their original description. The oldest, largest, and most diverse of these repositories...culture in the present day Cell culture is a widespread tool used in the fields of oncology, virology...problems surrounding cell culture over the last few decades some pharmacologists have tended to avoid cell ...International Cell Line Authentication Committee includes a database listing all cross contaminated or misidentified...comparable studies to be performed on a cell line even decades after it is initially isolated. Regrettably this...Master and Working stocks to work from. The future Attitudes are changing but yet more is needed to convince...

...plasmids are passed down through generations (Figure 1). These so called “plasmid addiction systems” require...than plasmids without this DNA. They eventually designated this fragment as the ccdB region (for “coupled... serves as a counterselectable marker in the destination vector. Successful cloning occurs when the ccdB...restriction endonucleases in plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...long-term experiments. They’ve also helped researchers design and optimize cloning methodologies using counterselection...

... done and we’re onto the next story. Figure 1: Addgene’s Agile Approach Sharing is caring There...released a major redesign of how search works on our site. For Addgenies, development includes everything from...experience and the overall soundness of the software design. Each of us brings slightly different cognitive...Addgene. Another advantage of Scrum is that it provides transparency to everyone involved about the status... on a new feature. We start with a story that describes the feature and what the criteria is for accepting...our users, we are able to make informed software design choices, resulting in software that helps our users...

...inclusion bodies and is folding properly. References 1. Angov, Evelina, et al. "Heterologous protein expression...features to consider when designing a full gene. A non-exhaustive list includes: Codon abundance relative...that their codons are more appropriate for the desired expression host. This is known as “codon optimization...as easy as it sounds. Even for relatively short peptides, there can be many possible ways to encode them...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...PubMed Central PMCID: PMC4794256. This review provides a great overview of codon usage bias 8. Tuller...

...drives our bioinformatics needs       Fig. 1: QC'ing data at Addgene is like standing under a Niagara...of data. In this two-part blog series, we will describe the importance of bioinformatics at Addgene and...each week. Our internal software was originally designed to help us work in large formats, such as 96-well...Thanks to the flexibility inherent in Nextflow’s design, we were free to use multiple programming languages...alone! The benefits of containers are they are designed to ‘contain,’ in a complete package, all of the... who or which computer ran the process. Another design choice we appreciated about Nextflow was how effortless...small with a print statement and tested that the desired data was available so we felt confident we were...

... end sequences (R and L ends).  Figure 1: (A) Components involved in each type of CRISPR-Transposon...human therapeutics (e.g. using Sleeping Beauty). Despite their diverse utility, DNA transposons have been...(V. cholerae INTEGRATE - hereafter VchINT) and encodes a Type I-F CRISPR-Cas system. The four major components...the same promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also...: Clone spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid...even very large genetic cargo, at one or multiple desired genomic loci. Many applications of these systems...

...mammalian expression from human elongation factor 1 alpha Constitutive  Tends to give consistent expression...insert into RNA. In practice, the term "promoter" describes the combination of the promoter (RNA polymerase...   Primarily used for RNA     transcript     Description Expression Additional considerations CMV General... phosphoglycerate kinase gene.  Constitutive  Widespread expression, but may vary by cell type. Tends ...promoters     Promoter     Primarily used for Description Expression Additional considerations T7 in ...identify the right tissue-specific promoters as described by researchers at the NIH here.  Note: A. Max...

... reached labs in over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image from... developed based on 100 structure-guided spike designs and has improved protein yields and stability. ...Plasmids deposited from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding...Liu Y, Chou C-W, Byrne PO, Hjorth CK, Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D, ...Finkelstein IJ, McLellan JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Science...

... of low-toxicity RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using...The hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. ...two pieces: a single protein, computationally described as a fusion of four Cas7 proteins and a putative...RNA editing methods — streamlining experimental design and its potential for clinical applications. Specifically...high-fidelity rate. Both Cas7-11 and PASTE open up multitudes of possibilities at the bench and potentially...