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We narrowed to 587 results for: cel.2

Showing: 281 - 300 of 587 results

  1. Hot Plasmids - January 2023

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    Blog Post
    ...is always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid...the presence of oxidizing agents. Adapted from Fig. 2 Campbell et al, Nature Methods   Find YFP plasmids...mCherry for FACS sorting of the weaponized T-cells (CAR-T cells) - and then delivering them to a tumor - ...effectively as standard CAR-T cells, but it reduces the off-target effects caused by T cell migration.  CAR expression...reducing the on-target off-tumor activity of CAR–T cells was recently devised by the Yingxiao Wang lab.  ...above can be made for expressing your favorite T-cell receptor, or perhaps any other tool that can benefit...  Wu Y, et. al. Control of the activity of CAR-T cells within tumours via focused ultrasound. Nat Biomed...

  2. 5 Great Apps for Lab Life

    Type
    Blog Post
    ... you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find an app that could...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...tools are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution...educational section. This section holds the Gibco Cell culture manual and basic instructional videos. The... Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England BioLabs...

  3. Evolution of Lab Techniques

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    Blog Post
    ...mutation is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new prenatal test for sickle-cell anemia." ...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically advanced... of the mutated β globin gene that causes sickle cell anemia. The mutation happens to destroy a sequence...conducted on a sample from an individual without sickle cell anemia, two DNA fragments are observed because the... site where MstII cuts. In a patient with sickle cell anemia the enzyme does not cut the DNA so only one...

  4. CRISPR 101: Off-Target Effects

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    Blog Post
    ...generate a knock-out cell line to perform downstream characterization of, isolating 2 or 3 distinct clones...sites would confound your experimental results and 2) perform targeted off-target measurements at those...experiments where only localization of the Cas occurs. 2 gRNA approach Off-targets that come from mutations...millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to sequence...trying to generate a knockout cell line, with the goal of isolating single cell clones to use for multiple...generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation arising.... a level of, say 5%, then the majority of single cell editing events will be on target. Thus, if you isolate...

  5. DAP arrays for multiplex gene editing

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    Blog Post
    ...disease, and type 2 diabetes, across four different types of human cells, including T cells and hematopoietic...(#186715) and hCtRNA_VT (#186716).   Fig. 2: Comparison of previous representative dual-gRNA architectures...to enable multiplex gene editing in a variety of cells and organisms. However, lengthy genetic payloads... over 50% averaging editing efficiency in human cells. MBE with DAP array also exhibited reduced Cas9-...hematopoietic stem and progenitor cells. Now with the DAP array, it is possible to use the state-of-art base editor... editing, and nuclease editing, in virtually any cell type or organisms of interest with only minimal ...

  6. Plasmids 101: Common Lab E. coli Strains

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    Blog Post
    ...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R ...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular...degraded, cell still can methylate DNA   hsdS(rk-,mk-) Unmethylated DNA not degraded, cell cannot methylate... gal Mutation in galactose metabolism pathway Cells cannot grow on galactose only gyrA, gyrA96 DNA...permease inactivated, lactose cannot be taken up by cell mcrA, mcrBC Inactivation of pathway that cleaves...gene that maintains the plasmids at ~250 copies per cell.  F- ∆lac169 rpoS(Am) robA1 creC510 hsdR514 endA...rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning and storage of unstable DNA and lentiviral...

  7. Synthetic Photobiology: Optogenetics for E. coli

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    ...Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication ...Levskaya A, Voigt CA. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010...Voigt CA, Ellington AD. Cell. 2009 Jun 26;137(7):1272-81. doi: 10.1016/j.cell.2009.04.048. PubMed. Find...with sophisticated gene expression controls. Most cell types do not respond to low-intensity light, so ...

  8. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ...same parent cells that represent the preexisting resistant subpopulation 2. Resistant cells among the separate...440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct evolutionary...resistant cancer cells exist prior to drug treatment. They infected a non-small cell lung cancer cell line containing... target cell population at low multiplicity of infection (MOI) such that each individual cell receives...researchers to individually label millions of mammalian cells through lentiviral infection and to monitor their...quantify the take rate of mouse xenografts after cell implantation. Their goal was to figure out how to...ClonTracer library to determine whether or not cancer cell populations resistant to drug treatments arise during...

  9. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ...parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each...activating caspase-3, and detected fluorescence within 2 to 5 hours, a time consistent with previous methods...the degradation of over 600 cellular components ultimately leading to cell fragmentation and death (Elmore...issues with tissue autofluorescence, cell heterogeneity, and/or cell position changes (To et al., 2016, ...apoptosis in HeLa cells, zebrafish embryos, and Drosophila. They treated HeLa cells expressing FlipGFP...cleavage by executioner caspases. Failure for cells to die or cells dying at the wrong time and/or place can...Apoptosis or “programmed cell death" plays a pivotal role in an array of biological processes including...

  10. Top Requested Lentivirus and AAV of 2016

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    Blog Post
    ... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...PMID: 24086725. PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher...expressed in cells that express cre recombinase. Typically, this feature is used to attain cell-specific ...tools so that researchers around the world can accelerate their work. After all, as we like to sayat Addgene...CRISPR applications and can be expressed from your cell line of choice after transduction with this virus...blast virus can be used to generate Cas9-expressing cell lines, which can then be used both for screens (...viral prep to create a single stable Cas9-expressing cell line and then transduce this line with lentiviruses...

  11. Twenty Years of Sharing: Expanding Accessibility

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    Blog Post
    ...China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor Partnership...distribution after we partnered with them (Figure 2). In others, the distributor helped solve an issue...temperature for the duration of their journey. Figure 2: Annual distribution of items to countries with a ...we are today. As part of our 20th anniversary celebration, we thought it only fitting to share some stories...

  12. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Blog Post
    ...the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, R6K, and ColE2...biotechnology 36.4 (2018): 352. PubMed PMID: 29553576. 2. Kittleson, Joshua T., Sherine Cheung, and JChristopher... engineer cells for specific purposes. Unfortunately, plasmid copy numbers vary within cell populations...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes.... When cells are dividing rapidly, multiple rounds of replication are initiated before the cell has divided...growth rate can all impact plasmid copy number in a cell. One can attempt to eliminate fluctuations due ... pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered against the effects...

  13. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    ...VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2; 3. The MS2...Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;156(5):935-49. doi: 10.1016/j.cell.2014.02.001. Epub 2014 Feb...Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct...screening for genes that, when turned on, make melanoma cells resistant to treatment with a BRAF inhibitor. (The...Addgene, enabling systematic investigation of the cellular consequences of repressing or inducing individual...Weissman and his colleagues demonstrated in a paper in Cell that they could typically achieve 90-99% knockdown...straightforward to apply the new library for use in various cell types and in live mice. Therapeutic uses in humans...

  14. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...check its subcellular location and solubility. If it’s membrane-bound or located in a subcellular compartment...reagent. Be aware that this approach requires excellent attention to detail, since you’ll end up with ...

  15. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...models or cells lines, including stem cells, without leaving behind any accessory DNA sequences [2], [3]....leaving behind only those cells with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D...transgenic cell lines? One of the major uses of the piggyBac™ system is to create stable transgenic cell lines...efficiency is likely to be cell line-dependent, and should be tested in the cell line of interest. Importantly...If the cell’s chromatin structure undergoes significant rearrangement (such as when a stem cell initiates...applications, including making transgenic cell lines and genetically-modified cell lines. Furthermore, the use of...

  16. Viral Vectors 101: Inverted Terminal Repeats

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    Blog Post
    ...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ...allow scientists to express a gene of interest in a cell or animal model. AAV plasmids contain several important...Integrity on rAAV8 Production Using the Baculovirus/Sf9 Cells System. Hum Gene Ther Methods, 28(5), 277–289. https...

  17. Targeted Mutagenesis with EvolvR

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    ...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD...EvolvR could also be a tool for lineage tracing cells that do not tolerate double-strand DNA breaks by...researchers to trace the progeny of a single starting cell. Additionally, multiplexing of guides allows EvolvR...

  18. History of CRISPR Cas - A tale of survival and evolution

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    Blog Post
    ...phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip..., accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/Cas moieties carry out these functions...devise new strategies to reproduce within a host cell. A few virus particles have learned to circumvent...

  19. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

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    Blog Post
    ...one injection step! The entire protocol takes about 2-3 weeks but requires less than eight hours worth of...for C. elegans genome modification. Figure 2: Schematic from Dickinson et al. (2015) illustrating...systems may prove effective in other species or in cell culture. In addition to CRISPR/Cas9 applications...

  20. R Bodies: Membrane-Rupturing Microscopic Tools

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    ...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies... can rupture E. coli spheroplasts (cells which have had their cell walls removed with lysozyme). This ...protein fusions allowed us to image R bodies inside cells, and these constructs may be useful to future scientists...applications. For example, you could use them to lyse cells (and surrounding membranes) in keeping with their...penetrate other barriers (like vasculature or extracellular matrices) in response to pH changes. Beyond...assemble and function, we might be able to endow cells with new ways to interact mechanically with their...
Showing: 281 - 300 of 587 results