We narrowed to 903 results for: nes

...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding...Homology Directed Repair (HDR) to knock in corrective genes. For this you would need to provide a DNA template...successfully developed and used AdV vectors to edit genes in several biological systems. It has been used ...plasmids at Addgene: pAdSh.PGK.Cas9 (expresses S. pyogenes Cas9 from the PGK promoter) and U6 promoter-driven...CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4:...

... (24-30°C). Brightness: Brightness is a measure of how bright is the emission. Brightness is calculated...contributed by Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how fluorescent proteins glow...the protein, divided by 1000. In many cases the brightness is compared to that of EGFP which is set as 1...proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be taken into account...

...namely that they can cause cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular... are, in most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet ... is (thankfully) gutted of many essential viral genes. How it is, exactly, that heaps of viral vector ...the only component that is not dependent on viral genes. It is derived from the host/producer cell membrane...full “life cycle” of expression of selfish, lazy genes of a non-living viral particle, and how we as dedicated... N. (1971). Noninfectious ARK mouse embryo cell lines in which each cell has the capacity to be activated...

...changing guidelines and regulations impacting grant management, we track grant opportunity deadlines, and ...organization to evaluate the organization’s grant readiness, identify grant opportunities, secure grant awards...schools, healthcare facilities and with private businesses. How do you get the expertise needed to enter...such as research, collaboration across multiple disciplines, writing, project management, and program evaluation...process or the whole process, including grant readiness, funding opportunity research, application writing...

...love with biology because of an image that was honestly quite boring. My Bio 101 professor displayed a...mammalian vectors are even less similar to bacterial ones. The mammalian analogue of bacterial transformation...uptake, but this approach requires compatible cell lines that express viral machinery. Alternatively, some...selection is useful for producing stable mammalian cell lines, but it isn’t common for simple transfection experiments...by transformation and transfection and left the business of transduction to our Viral Vectors 101 series...Resources on Addgene.org Collection of empty plasmid backbones ...

... transition Base change between purines (A ↔ G) or between pyrimidines (C ↔ T).   Base transversion ... ABE would allow it to act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the...this promising start, the team used rational mutagenesis to produce the adenine transversion editor AYBEv3...and ABEs, they could be useful for saturation mutagenesis, and they provide a strong proof of concept for...empty space. Over several rounds of rational mutagenesis, the team progressed to gGBEv6.3, which was capable...

...that recognize codons found in highly expressed genes (Emilsson and Kurland, 1990). Controlling gene expression...changing conditions - Organisms often need to express genes at different levels under different conditions. ...specific tRNA pools. For example, tRNAs used in genes encoding amino acid biosynthetic enzymes may be ... the problem of codon choice is to resynthesize genes in such a way that their codons are more appropriate...properly. You may be able to avoid getting your genes of interest codon optimized by ordering plasmids...use the same expression system for many different genes without having to create new constructs. However...

...activators and repressors to target genes. Repressing target genes using dCas9-based repressors Early ...repression or knockdown allow you to study essential genes whose knockout would cause cell lethality. The Krüppel...purpose. dCas9-KRAB plasmids for repressing target genes in a variety of species and cell types can be found...found on Addgene’s website. Activating target genes using dCas9-based activators The simplest of the dCas9...transcriptional activator. Plasmids for activating target genes in a variety of species and cell types can be found...

...engineering pipelines, we selected Nextflow, a workflow management system for scientific pipelines, including... existed who had created a shared resource of pipelines for genomic data. Thanks to the flexibility inherent...separate Nextflow process. In Nextflow, a process defines which command or script will run and can be thought... connect each subsequent process to the existing ones. Once our first process was working as expected,...check, we repeated our strategy and copied a few lines of our existing script into the section of the next...

...job it is to disagree with the group. They are the ones who ask for rationales behind ideas and decisions...fill out the form at any time and a member of our Business Development team would review it. This survey ...frequently observe in a scientific setting.  Powerlessness It’s hard to be creative when you don’t believe...ideas obviously worth pursuing, evaluate the “bad” ones. Bad ideas are easier to work with than nothing ... mistakes that kill your productivity. Harvard Business Review. Retrieved January 14, 2022, from https...exercise to spur innovation on your team. Harvard Business Review. Retrieved January 14, 2022, from https...

... the speaker and offer your business card (yes, you should have a business card; it’s a bit more informal...smaller talks within the bigger conference, especially ones that are more career-focused. After these talks,... will probably be willing to accept your CV or business card. Be warned, though – someone accepting your... formally apply for the position. This post on Business Insider nicely summarizes good job fair practice... Reading and Outside Resources http://www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014...

...in mouse embryos to generate gene modified mouse lines. Hopefully this post will help get your in vivo ...that later). Find Validated gRNAs to Target Mouse Genes To begin the in vitro screening process, you should... at the same time, and choose the most effective ones to increase your chances of gene modification in...a rapid acceleration in the production of mouse lines that could be used to study a wide variety of phenomena... et al. "Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes....generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering." Cell...

...Commercial Plasmid Prep Kit Amount 1000 Lanes 1000 Lanes 1000 Lanes Cost $200-$500  <$10 $20-30 Cost...commercial ladders cost between $200-$500 for 1000 lanes worth of ladder. The Penn State ladders are significantly...prep protocol, it cost less than $10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder. Using a ...

...I really brought to the table was empathy, a willingness to listen, and an ability to remember the right...amount of conflict is actually critical in the business world. Companies and organizations that avoid ...involved, and also for the labs, organizations, or businesses as a whole. When I’m attempting to resolve a ... actively runs away from it. That can mean a willingness to make yourself temporarily uncomfortable for...while giving people the tools to resolve necessary ones on their own is even better. The Thomas Kilmann ...conflict resolution based on various levels of assertiveness and cooperation: avoiding, competing, accommodating...

...productive discussion first defines the material resources needed, then determines how much those resources...possible, this even includes clocks, timers, and phones.  As for reagents and consumables (such as pipet...time to decide what resources to keep, and which ones to abandon. It’s critical to make these decisions...1. First make decisions based on experimental timelines and specific aims. 2. Then make decisions based...productive and manageable laboratory environments. His business skills enhance his passion for science. Follow...

...526) that (1) enables efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity... optimize conditions for a screen that assessed genes that regulate antigen-specific CD8+ T cell responses...screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8+ T cell responses to ... Ptpn2, Pdcd1, and Zc3h12a, demonstrating the robustness of the system and utility of the pRDA_526 vector...introduction of the model antigen, Ovalbumin, into cell lines to study antigen-specific responses. Find FITS plasmids...

...focused on choosing a lab, but almost all of these guidelines can apply to any workplace or job. It amazes ...will need is about you. It’s worth taking time to honestly evaluate your own style of work and communication...below, the first question to ask yourself is “which ones matter to me?”  and hone in on the factors that ...Overwork – including undue pressure, impossible deadlines, unnecessary disruptions Destabilization – including...author order PI lets lab members fight it out PI intervenes if there are interpersonal conflicts PI has no...can be very helpful in uncovering strengths and weaknesses of the options. Use this blog as a guide. Start...

...transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off, literally at the flip of...are able to react and learn. All of this requires genes to be regulated dynamically.” To understand how ... tools to manipulate the expression of specific genes in the brain in reversible fashion. And that’s exactly... of living mice both quickly and significantly. Genes under LITE control showed a rise in transcription...researchers applied the LITE system to 28 different genes to establish it as “a novel mode of optogenetic ...

...frozen bacteria, preventing damage to the cell membranes and keeping the cells alive. A glycerol stock ...

...and many plant scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity...than that from the commonly used 35S promoter. Transgenes are also efficiently inherited in both transgenic...expression. Sahoo et al. have tested the system with genes up to 4 kb in length and confirmed successful protein... assays as well as stable expression of foreign genes in plants." PLoS one 9.6 (2014): e98988. Pubmed ...Plant Synthetic Biology Quick, Versatile Plant Transgenesis with GreenGate Plasmids Interview: Nicola Patron...

...dictator or laissez-faire; theory or practice; business-like or artistic; tech, anti-tech or pragmatic...employers love PhD students to have: working to deadlines, writing up clear and attractive reports, being...students and postdocs go on to work in patent law, business development, teaching, etc. Church also encourages...science and engineering (STEM) expertise in law, business, journalism, entertainment, politics, K-12, etc... passion, hard work, collaboration, and staying honest with personal goals. Stay tuned for the follow ...

...explosion SUperNova," SunTag can help you turn up the brightness in your fluorescent imaging experiments. Fluorescent...managed to track run lengths of the motor protein kinesin across microtubules. Tanenbaum et al. then tested...to traditional fluorescent proteins Improved brightness and signal-to-noise ratio Less chance of phototoxicity...used to improve CRISPR-based activation of target genes, but we’ll save that application for another day...on Addgene.org Find Fluorescent Protein Empy Backbones Visit Our Fluorescent Protein Guide Page Find ...

...naturally occurring genes for cloning purposes (antibiotic resistance genes, toxin genes, and the sacB gene...gene of interest, but others will uptake plasmid backbones that re-ligated back on themselves. Therefore,...Plasmid cloning: Creativity with naturally occurring genes  At this point in the blog post, you may have realized...

...both introduce genes into an organism’s genome (see figure) and to disrupt endogenous genes at the site ...by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature Genetics... the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest...transposons rapidly generate stable transgenic cell lines. Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821...

...research project must have innovative yet attainable milestones that contribute to intellectual thought and discovery... and skills do you need to use to reach these milestones? Can you get assistance from people who have ...Think about what your personal goals are and set milestones along the way so that you don't get stuck doing...been developed. Can you think of a project that combines an exciting new method or approach with another... Passion fuels the drive to continue persuing milestones and working hard. Being patient, being selective...

...Now that we know all about antibiotic resistance genes, let’s consider another basic element of any plasmid...focused only on those found in bacteria. Some common ones you might see include ColE1, pMB1 (which comes in...20-40 CloDF13 (CDF) D Relaxed This table defines common cloning vectors, their copy number, ori,...copies of plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain...for single- and multiple-unnatural amino acid mutagenesis in Escherichia coli. Biochemistry. 2013 Mar 12...

...validated simultaneously in the brain and the intestines of mice. The result? A very cool tool to advance...polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics included...engineering road map of sorts, giving an idea of the stiffness the fibers would need to be, as well as other ...fiber implants. Due to the soft nature of the intestines and the requirement for digestive motility to...excited a class of enteroendocrine cells within the intestines known as neuropods. They observed increased vagus...

.... Yet, we cannot prosper in a world where our medicines, technologies, defense, and governing policies...more scientists in our leadership, and raising awareness regarding the pitfalls of accepting alternative...chromosomes. Montreal, QC, Canada 45.5017°N 73.5673°W – Vanessa Sung Hundreds of scientists and supporters in ...well as allies outside the scientific community. Vanessa Sung is currently a PhD student studying breast...restorative justice.  Follow her on twitter @sung_vanessa. Albany, NY, USA 42.6526°N 73.7562°W – Nicholas...denying science, from celebrities falsely claiming vaccines cause autism to politicians saying that climate...

...mice Transgenic mouse lines have also been engineered to express various opsin genes [7-9]. The advantage...other tool in a scientist’s vast toolbox, its effectiveness depends on how and why it’s used. There are ...behavioral output is sufficient to assess the effectiveness of your optogenetic manipulations (though a ...has genetically manipulated a number of the opsin genes to alter the light spectrum for activation and the...advantage of using these lines is, if one is crossed with an appropriate mouse line expressing a particular ...

...Cecchetelli Auxin Inducible Degradation (AID) uses plant hormones called auxins to conditionally and reversibly ...AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al., 2016).  ...protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that auxinole...targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires an NGG PAM to recognize... role in the transcriptional regulation of many genes. A number of groups have designed means of studying...

...Julio Ernesto Valdivia Silva, Universidad de Ingeniería y Tecnología (UTEC), Peru Julio Ernesto Valdivia...her current research, Sheen aims to identify new genes related to PZA-resistance by using different approaches...experience in the field of bacterial and fungal pathogenesis. Her research focuses on a collection of pathogenic...growth will then be measured to link the deleted genes to specific functions in biofilm formation. The...

...provide the Brunello (targeting human genes) and Brie (targeting mouse genes) pooled libraries from the labs...popular genome editing tool is its developers’ willingness to make their CRISPR technologies available to...pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular processes...80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to both perform...

...when choosing an individual fluorescent protein (brightness, photostability, and so on; see the previous ...use (particularly not the most recently published ones), they can be a good starting point. In many cases... used filter sets for multicolor imaging include ones designed for CFP, YFP, and RFP or the Sedat Quad...supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved folding...proteins with this filter set and have reported brightness measurements. Here, EGFP outperforms the improved...

...stable cell lines and are not used for general plasmid selection. Instead, reporter genes are often employed...within the cell for effective replication. Cell lines expressing the Epstein–Barr virus (EBV) nuclear ...48 h post transfection when creating stable cell lines. Looking for Mammalian Expression Plasmids? Check...Read More: Plasmids 101: Antibiotic Resistance Genes Plasmids 101: Origin of Replication All Plasmids...

... comic, when a bacterium uptakes a plasmid, the genes on the plasmid can give them different phenotypes...example are bacteriophages that attach to bacterial membranes and inject their genetic material into the cell... DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral and Adeno Associated...and AAV can be used to create both transient cell lines, where a gene of interest is expressed but not integrated... integrated into the genome and stable cell lines, where foreign DNA is incorporated into the cell’s genome...

...activated by an array of different ligands, including hormones, neurotransmitters, photons or odorants, to elicit...that there are approximately 950 different human genes encoding GPCRs (Takeda et al., 2002). G proteins...be used to detect G protein activation via bioluminescence resonance energy transfer 2 (BRET2). Check ..., GPCRs were first mutated via site-directed mutagenesis to bind non natural ligands. These non natural...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520...

...transcription of other relevant viral genes (including the ones listed previously). E3/E1B suppresses ...applications and aid in ease of use. Our lentiviral guide outlines these components.     Fig. 1 - Viral vector... AAV also requires some back up from adenovirus genes – E4, E2a, and VA – which are supplied on a helper...viral replication. More information on these viral genes is available in the adenovirus section below!   ...

...single-use” transgenic lines. Instead, researchers can generate and maintain transgenic lines that can be used...system that makes use of pre-existing transgenic lines, termed BLITZ (Biotin Labeling In Transgenic Zebrafish...dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system... 12-plex multiplex imaging system. Mixing the subclones within a single well reduced well-to-well variability...

... nucleic acid sequence of the antibody-encoding genes over time as the hybridoma cells divide. These changes...hybridoma cells before culturing or they can clone the genes from the hybridoma into a plasmid to create a recombinant...Figure 2: Changes to the heavy chain or light chain genes within a hybridoma impacts the specificity, avidity...Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, ...1999. 1, Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies. https://www.ncbi.nlm.nih.gov...

...target gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant... cAMP and represses pBad, decreasing promoter leakiness.   Other examples of chemically induced promoters... be compatible with your experimental system. Leakiness If you’re expressing a gene that may be toxic,...inducible promoters are also known for their low leakiness. Inducibility level How much do you want to induce...