We narrowed to 321 results for: POR C

... Since the bacteria are subsequently grown at 37 °C, the recombineering plasmid is not maintained. Recombination...the deletion. REPLACR-mutagenesis successfully incorporated a 60-nucleotide insertion; beyond this insertion...

...ti2learning.org. References 1. Dieterich, Daniela C., et al. "In situ visualization and dynamics of newly...designated work areas and come with health risks. Importantly, with this method it is not possible to follow...azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins instead of methionine...endogenous protein of interest that also had incorporated the AHA tag after pulse labeling of a determined...FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover of specific endogenous..."Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains." Proceedings...Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings of the National Academy...

...and References References Wang, I. X., Grunseich, C., Fox, J., et al., Human proteins that interact with... how these structures are generated, why it’s important for cells to sense their formation, and how clinicians...in turn lead to genome instability. Thus, it’s important to strike a balance between productive levels ...be delivered or anticipating this response are important considerations when using these reagents. RNA...recognized by RNase H which hydrolyzes the RNA portion, allowing the DNA strands to re-pair. The two dominating...stranded isn’t just for DNA. Untwisting RNA is important for therapeutic delivery, viruses, human disease...

...Methods Mol Biol, 2018. PubMed PMID: 29043675.Grefen C and MR Blatt. Biotechniques 2012. PubMed PMID: 23066669...2017). However, Acr inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument...versatility opens up the potential for spatial and temporal control in many genetic targeting and editing ...rapidly screen thousands of proteins for one that can report electrical activity via imaging. They took the ...response to light, offers a way to spatially and temporally control cellular processes in vivo. Many photoactive...

...proven in (A) Productize, (B) Roll out to market, and (C) Break even cycles. Work hard. Work fast. Be flexible...bacteriophages as bacterial therapeutics. This idea was incorporated into Novophage Therapeutics, Inc. in March 2009...potential conflicts that waste money and time. Most importantly, “the company does not become real until someone...Synthetic Biology? Read On! Synthetic Biology & the Importance of Assembly Standards Story of a SynBio Startup...

...binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling various... provides a useful yet low-pressure teaching opportunity for lab members or students new to protein expression...induce high levels of gene activation for both reporter and endogenous genes. It is highly specific in...

...specific genome editing. Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ, Aryee MJ...sequence flexibility indicates that gRNAs could be incorporated into these constructs without compromising cleavage...slightly higher mutation rates than they’d previously reported in protoplasts (15% and 9% of target sites.) How...

...targeting sequence of cytochrome c oxidase subunit VIII, so that they can be imported to the mitochondrial matrix...pCoofy series of plasmids contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP...N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator, and plant resistance... to report mitochondrial turnover Mitochondrial health, transport and turnover are important features...genes, reporters, or other markers into a target genome. They did this by selecting the important elements...usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer from Yan’s ...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to...

...Missinato, M. A., Onuoha, E., Perkins, L. A., Watkins, S. C., . . . Bruchez, M. P. (2016). A genetically targetable...photosensitizer fluorogenic dye.  MG-2I is the TAP portion of the complex and is derived from the malachite...

...Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth...of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse...

...sequence similarity to previously identified Cas13a-c orthologs, but they do include HEPN nuclease domains...localization signal (NLS). In a HEK293 mCherry reporter assay, CasRx and AdmCas13d produced 92% and 87%...

...python, R, perl, and ruby // This is a comment in C++ and Java Command line history: Tracking command...your methods section about the edit that was temporarily used. Just be sure that the original code remains...

...combination therapies.         References 1. Hyo-eun, C. Bhang, et al. "Studying clonal dynamics in response...leukemia. Bhang says the work highlighted the importance of developing more potent ABL1 allosteric inhibitors...

...References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O, Chambon P, and Ghyselinck NB. A directional...gene. Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...shown in Figure 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation...the two pairs of target sites. By flanking the reporter with one loxP site and one lox511 site in the ...

...antibody structure with important features labeled. V = Variable domain. L = Light chain. C = Constant domain...from another portion. For example, Fab fragments are molecules composed of just the Fab portion of the antibody...between the target and reporter. They have fewer non-specific binding opportunities due to the lack of an...region is the “tail” of the antibody and comprises portions of the constant regions of the two heavy chains...used in vivo.     Of course, the lack of an Fc portion means that most of the common anti-IgG secondary...it either needs to be directly conjugated to a reporter or you need a light-chain specific conjugated ...fairly easy to find these in a Fab fragment format. Reporter-conjugated anti-IgG Fab fragments make good secondaries...

...stimulation of dual virus-expressing CR neurons. (C) Duration of painful responses in the left and right...changes. In addition to supporting in-person conference attendance, the award now supports virtual conference...research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research ...third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study glutamate, a signaling molecule...in pMinDisplay (PDGFR anchoring domain), in electroporated in rat hippocampal cultured neurons, imaged...hydropersulfides modulate protein activity and have important roles during oxidative stress. To monitor this...

...manipulation. Common inbred strains include C57BL/6, Balb/c, and 129. Conversely, outbred strains were originally...gene incorporated into their genome. While transgenic mice have the new gene construct incorporated randomly...embryonic stem cells. Non-homologous recombination incorporates this DNA randomly into the mouse genome. In ...homologous recombination. A targeting vector is electroporated into embryonic stem cells and this will insert...vectors used to create knockout mice include a reporter gene, which replaces the targeted gene and provides...

...their monovalent nature. They are stable up to 80°C and can refold correctly after denaturing.  Designed...combinatorial libraries, and high specificity has been reported for a number of targets. Due to their size and...ain't broke, don't fix it!  That being said, it's important to know all your options when planning and running...

...doi.org/10.1038/nprot.2007.267 Stoeckius M, Hafemeister C, Stephenson W, et al (2017) Simultaneous epitope and...find yourself doing your own conjugation, it is important to keep in mind that you will need to confirm ...

...References and Resources References Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. A modular cloning...phenotype, translational output, etc., which can support a larger cellular engineering goal, several of ...

...islands (regions of the DNA with a high frequency of C and G nucleotides). The conversion of cytosine to ... versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27...The K9 reporter uses the HP1 chromodomain to recognise lysine 9 methylation, while the K27 reporter uses... methylation in living cells. Essentially this reporter construct consists of one substrate (the histone-derived...-K27 histone methylation reporter) positions on histone H3, which are known to be involved in repression...methylation of these sites was controlled, the reporter consistently showed significantly different FRET...states. Thus, this system is a reliable tool in reporting methylation status of these sites. This new technology...

... PubMed Central PMCID: PMC3622210. Schek N, Cooke C, Alwine JC. Definition of the upstream efficiency ...expression levels, the terminator also plays an important role in RNA processing and contributes to variability...polyadenylation. The addition of the poly(A) tail is important for stability of the mRNA, protection from degradation...degradation, and is integral to the nuclear export and translation processes as well.   Figure 2... non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and/or use...the Mason–Pfizer monkey virus constitutive RNA transport element lead to enhanced HIV-1 Gag expression ...

...procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can be used to confirm...Professor at Boston College. National reports stress the importance of providing authentic research experiences... is particularly interested in strategies to incorporate research into undergraduate courses and in communicating...

... Cas9 to generate the mutant cells, respectively (C). Mutant cells are screened (D) and "hits" are identified...post was updated on Aug 20, 2020. What genes are important in your phenotype of interest? Many scientists...not entirely known. Identifying which genes are important for a phenotype can lead to a wealth of additional...strong evidence that a particular gene is actually important for a given phenotype. Each hit from the screen...

...//doi.org/10.1016/s0091-679x(08)60255-1  Manning, C. F., Bundros, A. M., & Trimmer, J. S. (2012). Benefits... reactive ones. And make sure to check for any reported cross-reactivity when planning your experiment...Why Choosing the Right Secondary Is of Primary Importance. In J. Sturtevant (Ed.), PLoS ONE (Vol. 7, Issue...

...utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which result in the internalization...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane...and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target effects can ...through a process known as CRISPR-EZ (CRISPR RNP Electroporation of Zygotes) (Chen et al. 2016). For CRISPR ...CRISPR experiments that involve HDR, electroporation can be combined with cell-type specific reagents in a ...a technique known as nucleofection, which forms pores in the nuclear membrane, allowing for entry of a... account with these types of experiments. One important consideration is that the Cas9-gRNA RNP will be...

...media, reducing the incubation temperature (e.g., 30 °C), and limiting your growth time (e.g., 16-18 hours...individual libraries can vary significantly, it’s important that we have an amplification protocol developed...and the desired coverage for the library (How important is it that every single plasmid in your library...protocol. Created with BioRender.com.   The most important factor to consider is your desired library coverage...not millions, of plasmids, they are generally electroporated into electrocompetent cells to ensure the highest...chemical transformations is less than that of electroporation, but an advantage is that chemical transformations...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...

...the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist ...a fluorescent emission wavelength in the green portion of the visible spectrum (hence the name), which...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...solvent. This tightly-packed structure explains the importance of the entire GFP protein, which is almost completely...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...protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest,...

... targeting mCherry reporter (mChe), with co-incubation with assist peptide (AP). C) Workflow for Cas-RNP-PAGE...difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection.... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized by pre-existing antibodies...

...e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha N, Gil S, Wang H, Papayannopoulou T, Lieber...pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR activity... way to make those genome edits. But, just as important is a way to flip the switch and turn off CRISPR...The ability to turn off CRISPR activity is an important step towards CRISPR-based therapies and for more...

...     References and Resources References Shen C-H (2019) Amplification of Nucleic Acids. In: Diagnostic... a fluorescent signal. The signal increases proportionally to the amount of product produced each cycle...multiple starting DNA templates in one sample. It is important to note however, that multiplex PCR is more complicated...

...assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011...have that conversation out in the open is really important. Addgene: Is there anything else you’d like to...biology is massively underfunded considering how important it is. I would really like to reach all those ...

...4. Mittereder, Nanette, Keith L. March, and Bruce C. Trapnell. "Evaluation of the concentration and bioactivity... quickly due to immune response against AdV. Importantly, recombination-competent adenovirus (RCA) can...

...due to camelid high body temperature (37.2 - 39.2°C). The Seeger lab found a way around this by using ... Signaling and Protein Purification RANbodies: Reporter Nanobody Fusions Plasmids 101: Secondary Nanobody...

... where they could bind to intracellular epitopes. C) Permeabilizing with a non-ionic detergent, like Triton... for antibodies that bind to the extracellular portion of a protein. B) Permeabilizing with the selective...

...AAV11 is reported to target skeletal muscle, kidney, spleen, lung, heart, stomach, and brain. C) Recent... available with T-Sapphire, EGFP, or mScarlet reporters, so you can choose a color compatible with the...and AAV11 tropism and applications. A) AAV6 is reported to have broad tissue tropism, including skeletal...poses challenges, from non-selective anterograde transport to a lack of genetic control. ATLAS (anterograde...the Cre recombinase payload activates a floxed reporter gene (genetically encoded or delivered separately...

...20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia R, Marillonnet S. A one pot, one step, precision...produce complex fluorescent reporter constructs. These fluorescent reporters can be used to detect structure...), the same cloning procedure can be used to incorporate each element into the final construct. This strategy...protein sequences to develop diverse fluorescent reporters is a useful application of high-throughput cloning...sequences), so enzyme choice is critical. May not support in-frame fusions of coding sequences Sladitschek...

...Moreno-Mateos, M. A., Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed...typically require an exact DNA sequence to be incorporated at a precise genomic location, with little to...template for repair, which will result in the incorporation of your sequence, a.k.a a knock-in! So how does...higher HR frequencies than quiescent cells. It is important to know that HR is often mutated in cancer cells...molecule is used as the template, you need to incorporate homologous sequence to the right and/or left ...

...be used for promoter swapping and generating N and C terminally tagged fusion proteins with fluorescent...analyzable transgenic in hand, so it is pretty important to get this process right the first time. I’ll...to transform when they begin flowering. It is important to perform the transformation not long after flowering...cut off the primary inflorescence (the flowering portion of the plant) near the base of the stem. This buys...s time to select for transformants. It’s very important to select multiple T1 plants from each transformation...

..., Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl G, Tebas P, June CH, Humeau...transduced rat neurons in vivo with beta-gal and other reporters, and they showed that expression persisted for...Figure 2). The HIV genes that do remain are very important for viral production: Gag (structural precursor...activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A different viral envelope protein...

...reducing the chance of an opportunistic infection like for organisms like C. difficile. In 2014, Citorik...species, or even an antibiotic resistance gene. Importantly, native, non-pathogenic bacteria remain to help...colonization model, phagemid treatment decreased the proportion of virulent S. aureus from 50% to 11% in just...

...Central PMCID: PMC6247916. Felker, A., and C. Mosimann. "Contemporary zebrafish transgenesis with Tol2 and ...recent lineage tracing approaches that led to important insights in developmental biology. Microscopy-...pathway regulated by one-eyed pinhead, incorrect proportions of otherwise normal cell types are produced, ...

...red, green, or blue (RGB) fluorescent protein (4a-c). When high doses of this viral cocktail are injected...gut, and other organs to the CNS, are also an important target for gene delivery, especially for the study...3293995. Additional Resources on the Addgene Blog Important considerations when using AAVs Learn more about...

...perfect. Good luck! References 1. Haery, Leila, Ryan C. Thompson, and Thomas D. Gilmore. "Histone acetyltransferases... member of a class of proteins was mutated, as reported in various studies. We made a table that listed...for each protein we listed all the studies that reported mutations in that protein (including how frequently...

... mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  As the name implies, this promoter... to therapeutic uses, and in those cases it's important to identify the right tissue-specific promoters...Posts Learn about Mammalian Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org...

...Zou, Y., Mason, M.G., Wang, Y., Wee, E.J., Turni, C., Blackall, P.J., Trau, M., & Botella, J.R. (2017)...Purification and Gel Extraction Kits. Scientific Reports. https://doi.org/10.1038/s41598-018-30316-w  Zou...

...This is possible because there are 4 bases (A, T, C, and G) and 3 positions in each codon. There are therefore...isoacceptors induced by amino‐acid starvation." EMBO reports 6.2 (2005): 151-157. PubMed PMID: 15678157. PubMed...

...pdb-top63. PubMed PMID: 20150100. 4. Shaner, Nathan C., George H. Patterson, and Michael W. Davidson. "Advances... and fluorescent proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal...

...either the PAM (changing one of the G’s to an A,T or C – but avoid NAG) or guide RNA target (“seed”) sequence...it is incorporated. We were surprised to see how rapidly the efficiency of mutation incorporation falls...contains the intended sequence changes to be incorporated into the edited genome. In mammalian cells such...these two strategies allows you to specifically incorporate homo- or heterozygous mutations. 1. Increasing...therefore strongly encourage any researcher to incorporate CRISPR/Cas-blocking mutations at both targeted... our frustration) in many cases we only saw incorporation of the blocking mutation. Despite the fact that... away from the cut site were not efficiently incorporated during HDR. We decided to systematically characterize...

...PubMed PMID: 7915669. Dominguez, Roberto, and Kenneth C. Holmes. "Actin structure and function." (2011). PubMed...genes important for mesoderm differentiation Now, let's turn tinman and bagpipe, two genes important for...protein Groucho, a transcriptional co-repressor important in development. Groucho family proteins are found...Dana Farber Cancer Institute! Sonic Hedgehog, an important regulator of embryonic development The largest...video game character but rather to an extremely important gene in animal development. To understand where... Tinman, 2016). Bagpipe on the other hand, is important in the formation of the visceral mesoderm which...