We narrowed to 903 results for: Nes

...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...

...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...endonucleases. A given prokaryote typically has genes encoding one or a few restriction modification systems...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...coli strain and will therefore have methylated adenines in any GATC sequences found in the plasmid. When...Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org...

...TRIM21 toxicity, Clift et al. created stable cell lines overexpressing TRIM21, finding that TRIM21 overexpression...contain changes in expression of TRIM21-associated genes, indicating that TRIM21 overexpression does not ... within 4 hr of treatment. In some primary cell lines, endogenous TRIM21 is sufficient for TRIM-Away, ...Recruitment for Epigenome editing by Cas9 (FIRE–Cas9), combines these approaches to enable rapid and reversible...that can independently control expression of your genes of interest. These transcription factors consist...

... al. "High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities." Cell...et al., 2014; Konermann et al., 2014) individual genes for studies interrogating their functions. The continual...list of effective gRNA sequences targeting your genes of interest. Thanks to the tremendous efforts made...Hongming, et al. "A CRISPR-based screen identifies genes essential for West-Nile-virus-induced cell death...

...into account (Fig 1a). The laboratory of Karmella Haynes at Emory recently contributed a set of plasmids...week. Taking advantage of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes an elegant...chromatin within a nucleus -> DNA compaction around histones -> Examples of histone-modifying enzymes and reader-effector...reader-effector proteins. b) Golden Gate assembly (backbones available at Addgene) with examples of potential... proteins quickly and accurately.   Priode and Haynes, protocols.io. 2021. dx.doi.org/10.17504/protocols.io.brgcm3sw...

...can now precisely control where and when certain genes are expressed to investigate their role in a specific...would behave very differently (distracted) from the ones tested at the beginning. Variability source Standardization...manipulation can also reduce expression levels of certain genes. A popular method uses Cre recombinase on transgenic...vectors are superior packaging vectors for larger genes - they can hold up to 8 kb, while AAV can only carry...Timing: High expression levels of the viral vector trangenes typically occur two weeks after injection. Therefore...

...engage broadly with diverse communities, to raise awareness about where science is and where it is going, ...Harvard Medical School, has a mission of raising awareness and starting conversations about both the benefits...participation in new precision medicine research. A willingness to confront these tough issues is essential for... of individuals from diverse backgrounds and disciplines, and everyone on staff takes on a variety of ...of their families. Having a broad interest and awareness of issues outside of your field of research will...

...lentiviral) generation The first step in delivering genes via lentivirus is deciding what packaging system...VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer gene...) while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how... can affect transfection efficiency. For some transgenes, transfection optimization simply is not enough...sufficient titer; this is often the case with large genes. This is due to inefficient packaging of large inserts...

...additional safety benefits: they lack all viral genes, including those that are responsible for integration...significant clinical efficacy against a form of blindness known as Leber’s congenital amurosis (LCA). This...RPE65. It is characterized by childhood onset of blindness and was thought to be untreatable until 2008. ...Furthermore, deeper understanding of disease pathogenesis will enable robust therapeutic outcomes. In ...decline in vision with gene therapy in childhood blindness." New England Journal of Medicine 372.20 (2015...

...presenting, it’s no wonder it was attended by over 300 Chinese scientists eager to hear current findings in nucleic...where one can shop for components of traditional Chinese medicine. There were hundreds of stalls selling...Lowe talked about his work on the 500+ human tRNA genes and, until now, we've overlooked regulatory fragments...may be involved in preventing silencing of self-genes. Dr. Mello presented a hypothesis that these pathways...are engineered to replace the drug resistantance genes in the bacteria thus making the bacteria sensitive...

... to summarize these principles it would be “assertiveness.” 7 strategies to become more assertive & resolve...resolve conflicts What Is Assertiveness? An assertive person is able to communicate their ideas confidently...your action item list and set some approximate timelines. Be sure that you keep your commitments, and if... you to follow through on your commitments. Assertiveness is a skill that will help you to resolve both.... An excellent resource to learn more about assertiveness skills is Dale Carnegie’s book “How To Win Friends...

...containing plasmids differing in their resistance genes, as one does not need to prepare separate batches...

... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...changes under magnetic field. Such changes could be harnessed or enhanced for future MagLOV-based tools, just... Ingaramo, M. (2024). Magnetic control of the brightness of fluorescent proteins. Zenodo. https://doi....., & Kusamoto, T. (2023) Single-Molecule Magnetoluminescence from a Spatially Confined Persistent Diradical...

...Origins of protein tags For about as long as the disciplines have existed, cell and molecular biologists have...developing generic strategies for detecting cloned genes (Munro and Pelham, 1984). The strategy they landed...1985). Myc was one of the earliest identified oncogenes and was a target of intense research. Evan, et...explore Myc’s role in cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding...tagging to detect proteins expressed from cloned genes: deletion mapping functional domains of Drosophila...

...loci targeted One Many Sample type(s) Cell lines Cell lines, primary cells Table 1: Key differences...multiple targets because different CAPTURE cell lines will have varying levels of dCas9 and gRNA expression...components eliminates the need to create stable cell lines and allows CAPTURE 2.0 to be used with primary cells...like to interact with, and how many promoters or genes each enhancer is interacting with. Super enhancers...

...SB_loop, SB_convex. These scaffolds contain serines and threonines at positions that can be randomized for...epitopes that are hard for antibodies to get to, like ones that are hidden in the molecular crevices proteins...Developing antibodies against proteins embedded in membranes is tricky because they have fewer hydrophilic ...diverse pool of sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger...

...a chance of seeing a phenotype since additional genes that can do the same function are less likely to...Essentially, the X, second, and third chromosomes are the ones you’ll work with. If YFG is somehow on the Y or ...Geneticists need a method for maintaining stable lines of genetic mutants, including in situations where...lose track of your mutation. To maintain stable lines of flies carrying a specific mutation, Drosophila...Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective. Their Snapdragon tool helps...

...contacts that bring apposing membranes together and can wrap the membranes into stacked structures including...every lab in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to...tubules can be grossly distorted into dense stacked membranes termed Organized Smooth Endoplasmic Reticulum ..., M.V., et al., Fluorescent proteins from nonbioluminescent Anthozoa species. Nat Biotechnol, 1999. 17...

...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...action of the most common antibiotic resistance genes and which applications are best (or worst) for each...plasmid of interest, preventing this issue. The genes and their antibiotics   Ampicillin Ampicillin, often...antibiotic due to stable integration of resistance genes in their genomes. If you need a plasmid that can... Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage...

...breast and ovarian lines commonly used in lab. It is recommended you check your cell lines for HR-related ...covered in this blog, with tips and tricks for harnessing the homology-directed repair pathway, designing...surprise then that these edits take more planning and finesse then their knock-out counterparts. To kick off ...experiment today! Get ready to identify knock-in clones by reading up on validating genome edits to find...