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We narrowed to 845 results for: ELL

Showing: 321 - 340 of 845 results

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

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ... 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... to stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold...fold (for p15a, R6K, and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro...

  2. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ... protein (CcdA) that protects the cell against the toxic CcdB. Cells that lose ccdA through the loss of...disrupted allowing cells expressing the recombinant plasmid to propagate. Any cells that contained non-recombinant...Assembly Cloning and Gateway® cloning technologies, as well as several others. Each method is unique and relies...which is responsible for plasmid maintenance during cell division. ccdB codes for the toxic protein (CcdB...broken double stranded DNA and ultimately causing cell death. ccdA, another gene found in the ccd operon...Gateway takes advantage of the same principle that cells will not propagate while expressing the gene. Briefly...can one work with a plasmid/gene that kills the cells expressing it? The answer lies in special strains...

  3. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    Published
    April 22, 2015, 2:06 p.m.
    ...through programming cells. In the context of therapeutics, if you want to program a cell to identify and ...translational activity a cell has and control how much it taxes the resources of a cell. The objective is to... to the regulatory networks within E. coli host cells. Image from Nielson and Voigt (2014). ...within the wood. There are different types of cellulose fibers, pattern, shape. All of that is controlled...and target a malignant cell, then you’d need the ability to turn on genes at different times. It requires...being used. One thing is that Cas9 is toxic. When cells carry CRISPR circuitry, it can slow their growth...combination of parts in Addgene along with the software to tell you which parts are required to get the function...

  4. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...specific cell types or entire organs in planarians. For example, the gene Piwi labels neoblast cells and the...techniques to alter gene expression or to label specific cells, but there are too few resources to teach students... required for both WISH and RNAi experiments, as well as how to perform worm amputations. They also created... in tissue. This is useful for labeling specific cell types and for viewing how gene expression changes...modified nucleotides (for example, Digoxigenin-labelled Uracil) that will bind to complementary mRNA within...Williams, Eric J. Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado, The...

  5. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...Davidson AR, Maxwell KL, Doudna JA (2017) A Broad-Spectrum Inhibitor of CRISPR-Cas9. Cell 170:1224-1233...Sontheimer EJ, Maxwell KL, Davidson AR (2016) Naturally Occurring Off-Switches for CRISPR-Cas9. Cell 167:1829...concern is that the longer Cas remains active in a cell, the greater chances there are for off-target edits...Streptococcus pyogenes Cas9 (SpyCas9) is the most used and well studied CRISPR system, N. meningitidis can also ...within the first six hours. When added to human cells six hours after the introduction of Cas9 RNPs, AcrIIA4...1233.e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha N, Gil S, Wang H, Papayannopoulou T,...CRISPR/Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development...

  6. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...fluorescent proteins available. Fortunately, several excellent reviews have collated data from various papers... monitoring sensitized emission in living plant cells." (2013). PubMed PMID: 24194740. PubMed Central ...source of error in suboptimal or undetectable FRET, well-characterized FRET reference standards can be used... alters the FRET efficiency. Detecting certain cellular changes with an intermolecular probe is often ...Miyawaki et al., 1997), was designed to measure intracellular calcium and published in 1997. Since that time...of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments can probe...concentration or activity of small molecules, detect cellular processes and signaling cascades, quantify mechanical...

  7. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...producer cell line, such as HEK-293T, production of viral particles, and transduction of a target cell line...best to be well-rested and alert the day of the procedure. Mice move much more quickly than cell cultures...become a regular part of basic biological research as well as clinical therapy. These biological tools are ...re derived from viruses that can infect people, cells, and animals. Some of these viruses are completely...best practices for handling lab-grown viruses as well as potential risks and how to minimize them. Built-in...While the virus can still enter (and thus infect) cells, this safety feature prevents the virus from replicating...replicating and spreading in non-target cells, including yourself as the researcher.   Figure 1:...

  8. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...purpose and mechanism of polyadenylation vary across cell types, but polyadenylation generally serves to promote...naturally occurring terminators, such as T7 and rrnB, as well as engineered high-efficiency terminators such as... thought that poly(A)s are used to control the cellular concentration of regulatory RNAs and may additionally...additionally act as a quality control mechanism to rid the cell of mis-folded RNAs. Eukaryotic termination and polyadenylation...transcribes tRNA and other short RNAs. Although not as well studied as prokaryotic termination, the basic processes... the nuclear export and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation...

  9. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface receptor...antigenic drift.    Starr et al Cell. 2020. https://doi.org/10.1016/j.cell.2020.08.012  Starr et al Science...Library generation and sequencing. From Starr et al Cell., 2020. (B) Mapping antibody-escape mutations with... SARS-CoV-2 spike RBD is displayed on the yeast cell’s surface. The Bloom lab used the library to identify... candidates were validated by IP-MS using HEK293 cell lysate where the antigens would be in their native...immunoprecipitate the entire complex in three different cell lines. The Gräslund lab plans to continue their ...protection in yeast bioproduction pathways By Gabrielle Clouse Claudia Vickers’ lab has created a novel...

  10. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ... primary cells. The authors used this platform to assay dozens of GPCRs across many cell types in...biosensor platform. Cell, 187(6), 1527–1546.e25. doi: https://doi.org/10.1016/j.cell.2024.01.028. New...Prime editing (PE), initially developed in human cells, has been uniquely challenging to apply in dicotyledonous...labeling approach using these constructs in HEK 293 cells (Smith et al., 2024). They then used single-molecule...constructs for tagging the kinesins KIF5A and KIF5B as well as backbones for generating other SNAPf-, CLIPf-...CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only for...recordings of spontaneous activity in human stem cell-derived neurons expressing ASAP5-Kv. C) Cultured...

  11. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...Fluorescent Protein (GFP), a protein from the jellyfish Aequorea Victoria. Since that discovery, the family... localization of that biomolecule inside living cells. FPs classified by the emission color (emission ... the variants blue FP (BFP), cyan FP (CFP), and yellow FP (YFP) were derived. For a breakdown of GFP, ...467 nm Cyan 474 - 492 nm Green 499 - 519 nm Yellow 524 - 538 nm Orange 559 - 572 nm Red 574 - .... Therefore, these can be used as “timers” for cellular processes following their activation. The four... longer-wavelength filters (e.g. GFP also emits yellow light). Note that some combinations of FPs can ...Guest Blogger! Gal Haimovich, PhD, is a research fellow in the lab of Prof. Robert Singer at Albert Einstein...

  12. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    Published
    May 7, 2019, 12:39 p.m.
    ... Ba/F3 and MCF10A cellular models. Ba/F3 and MCF10A are growth factor dependent cell lines and any mutations...attempting to characterize mutations in a pooled cell population, as mutations with strong phenotypic ...corresponding wild-type genes were constructed as well. They called this library the CTD2 FASMIC Plasmid...functional groups, based on how each one impacted cell viability (see figure). Most of the mutations were...to be “neutral,” having no significant effect on cell viability compared to a wild-type control. Approximately... the mutations, however, significantly increased cell viability, and these “activators” may be the direct...represented in this collection along with hundreds of less well-studied candidates. The possibilities are vast. ...

  13. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    Published
    Aug. 4, 2015, 2:30 p.m.
    ... are you’ve used a GFP derivative to conduct subcellular localization studies or make a reporter construct...shown to work in plant, Drosophila and mammalian cells, and it may find wide use in biological research...turnover rate. In many cases, however, a GFP-tagged cell line or organism has already been made, and it would...made by accident. After exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. ...excitation at 561 nm. This imaging process worked well with the common GFP variants EGFP, mGFP4-T, and ...spontaneous conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White circles...this technique may not be appropriate for high-pH cellular compartments. Sattarzadeh et al. have shown that...

  14. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ...Funded by the NSF (with Co-PIs Jessica Bell, Lisa Gentile, Ellis Bell, and Amy Springer), this group now ...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...identifying new genes required for stem and progenitor cell maintenance in planarians in Developmental Biology...diversity and inclusion in STEM disciplines (Bangera & Brownell, 2014; Duboue et al., 2022), as they increase ... his teaching style, with the former offering excellent training in improvisation and the latter making...CURE trainee goes on to win a Nobel!   Jason Pellettieri is Professor and Chair of the Biology Department...References and Resources References Bangera, G., Brownell, S. 2014. Course-based undergraduate research ...

  15. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    Published
    May 12, 2015, 1:32 p.m.
    ...functional in cultured Chinese hamster ovary (CHO) cells, as well as in cortical neurons in layer 2/3 of the ...recombination varies for different constructs and cell types. Second, Cre may induce recombination at pseudo...standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding domain-fusion...activity and specificity with low cross-reactivity, as well as low toxicity. FLP/FRT recombinases (from Saccharomyces...recombination efficiency. When expressed in neuronal cells, each line showed broad recombinase-dependent RFP...these recombinases were also active in mammalian cells. In experiments, B3 and KD were shown to be functional... damage induced by Cre recombinase in mammalian cells. Loonstra A, Voojis M, Beverloo HB, Al Allak B, ...

  16. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    Published
    March 27, 2014, 5:14 p.m.
    ...optical labeling and tracking of proteins, organelles, and living cells in a spatiotemporal manner, which is...eight millimeters – not single cells (yet) but small clusters of cells. Looking ahead What’s next, you...fluorescent proteins to observe what’s happening inside cells for at least a couple of decades. Green is the classic... were lots of fluorescent proteins made from jellyfishes and corals, but all of them fluoresce outside... vivo. “One of the reasons we can’t localize one cell at a depth of one centimeter with fluorescence is...on the other hand, are larger than the size of a cell, so they scatter much less. As a result, ultrasound...brighter, more photostable, smaller, and less toxic to cells. They’d like to make near-infrared fluorescent proteins...

  17. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    Published
    Feb. 13, 2018, 2:51 p.m.
    ...when working with cell culture, because it is easy to harvest large quantities of cells. However, when working...numbers of cells or when a high degree of signal to noise is desired. It also provides an excellent additional... been widely applied in many model organisms and cell lines. Although ChIP-seq is a relatively simple ...typically requires a large amount of input material, cells or tissue, to produce a strong enough signal over...protein of interest. First, nuclei from tissue or cell culture are isolated using lectin-coated magnetic...the usage of a strong detergent to permeabilize cells rather than to rely on nuclei extraction. They report...technique, specifically in terms of initial steps of cell or tissue preparation.   All in all, CUT&RUN offers...

  18. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ... Guide RNA and Target DNA. Cell 156:935–949. https://doi.org/10.1016/j.cell.2014.02.001  Ran FA, Cong ... Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell.2020.03.023 Additional Resources...acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency ...package each piece into an individual AAV. When a cell is transduced by both AAVs, the full length gene...SpCas9 which had modest editing rates in HEK-293T cells when compared to the full-length SpCas9. In 2016... DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F (2017) RNA editing with CRISPR-Cas13... allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: ....

  19. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...low-frequency cancer mutations from cell-free DNA fragments as well as health-related single nucleotide...Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Additional resources...similar types of HPV, HPV16 and HPV18, from human cells, at atomolar levels, within one hour. DETECTR thus...science.aaf5573 Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, Miao X, Streithorst JA, Granados ...10.1126/science.aam9321 Gootenberg JS, Abudayyeh OO, Kellner MJ, Joung J, Collins JJ, Zhang F (2018) Multiplexed...Gootenberg JS, Abudayyeh OO, Metsky HC, Durbin AF, Kellner MJ, Tan AL, Paul LM, Parham LA, Garcia KF, Barnes...of Diverse Class 2 CRISPR-Cas Systems. Molecular Cell 60:385–397 . https://doi.org/10.1016/j.molcel.2015.10.008...

  20. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    Published
    Dec. 13, 2016, 3:30 p.m.
    ...if the cell dies from injury, this protein leaks out into the cytoplasm, migrates out of the cell, and ...Keytruda) causes T-cells to more easily recognize and cause the destruction of cancer cells. Pembrolizumab...cause inflammation, which can cause mesothelial cells to mutate and ultimately form cancerous tumors.  ...resection), systemic chemotherapy to kill cancerous cells that may have spread beyond the point of origin,...diagnosed so late that these treatments do not work very well. The best option in such instances is palliative...successful in killing many (if not all) of the cancer cells that may be left behind after surgery, while being...and binds cells that can begin an immune response. This important biomarker can be used to distinguish between...
Showing: 321 - 340 of 845 results