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We narrowed to 845 results for: ELL

Showing: 141 - 160 of 845 results

  1. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...improved cell-free system to examine biosynthetic pathways A new tool for enriching ABE-targeted cells The ...Improved cell-free transcription/translation system to examine biosynthetic pathways  By Gabrielle Clouse...interest, and cell extract.    Graphical Abstract: Schematic representation of cell-free system,...successfully reconstituted in this cell-free system. In summary, this cell free system allows for a relatively...   New tool for enrichment of ABE-targeted cells By Ellie Adams Single base modifications through ABE...multiplexed and can be used in human pluripotent stem cells, a cell type that has been difficult to modify in the...Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems) have been...

  2. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity....*Pro-Tip*: The cells will become completely confluent and the media will turn yellow. Do NOT change the...more HEK293 cells and produce more rAdV to reach higher titers. Successfully infected cells will become...large transgenes, transduce quiescent and dividing cells, and do not integrate into the host’s genome. The...administration, which results in the death of transduced cells and loss of transgene expression (Interestingly,...gene delivery: Potential applications for gene and cell-based therapies in the new era of personalized medicine...can be generated if the E1 gene from the packaging cell line is transferred into the AdV by recombination...

  3. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    Published
    Nov. 7, 2019, 2:18 p.m.
    ...including Trends in Cell Biology, Cell, Molecular Cell, Developmental Cell, and Cell Stem Cell.  She was the...inception and launch of other Cell Press journals, including Cell Metabolism, Cell Reports and Joule. In September...Deborah Sweet from Cell Press. Deborah Sweet is the Vice President of Editorial at Cell Press.  She trained...the founding Editor-in-Chief of Cell Stem Cell, a role she held for 10 years, and played a major role in...by Deborah Sweet, Vice President of Editorial at Cell Press. Almost everyone who works in a lab struggles...and making sure you do what you can to help your fellow researchers advance science overall. Communication...communication of science very seriously. That’s why at Cell Press we developed STAR Methods to improve the structure...

  4. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ... brain, robust single-cell labeling is necessary to distinguish individual cells. The Brainbow-1.0 construct... May 5. PubMed. Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish... both of Harvard’s Department of Molecular and Cellular Biology, harnessed these attributes of Cre-lox... 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing in other systems, such...Brainbow highlights the trend in biology towards single-cell resolution imaging. Sanes and Lichtman endeavored...contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered cyan). Without Cre...caused partially by fluorophore photoinstability, as well as the tendency of fluorophores to aggregate in ...

  5. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...specific cell type rely on transgenic models that express recombinases like Cre or Flp in their cells of interest...entry into the cell (Dudek, AM. 2018).  If the serotype you are using transduces cells that you are not... an ATG start codon to prevent transcription in cells where the recombinase was not present (Kuhlman, ...recombination that can lead to leaky expression in cells that do not express Cre or Flp. Here at Addgene ...recombined/functional transgene that can be expressed in cells where Cre is absent. Additionally, we have experienced.../Flex vectors and leaky expression can occur in cells that don’t express Cre or Flp, what can we do about...volume needed to achieve adequate expression in your cells of interest.  The AAV serotype used can exhibit ...

  6. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    Published
    Oct. 9, 2018, 12:21 p.m.
    ...PFs need to tell and sell their stories to get jobs. PFs (and mentors) can never be too well practiced ...doing a postdoctoral fellowship, what are you signing up for and how can you do it well? Below, I briefly...exciting story I can tell with my data?  Can I make better slides that more effectively tell a story? Generally...Hughes Medical Institute. Today's postdoctoral fellows (PFs) face a number of challenges ranging from ...briefly describe some tips for a successful postdoc fellowship. 1. Reverse engineer success On Day 1 of your...preparation) A history of successful funding (fellowship grants) A project with sufficient preliminary...need to be there? A typical biomedical postdoc fellowship lasts five years. Most universities and institutes...

  7. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ... cultured cells as well as more difficult conditions such as model systems and primary cells. References...when trying to screen a limited number of cells like primary cells or in vivo. Recently, the labs of David... human CRISPR libraries. Brunello: human CRISPRko sgRNA library The Brunello library consists of 77,441...criterion for probing a limited number of cells. Get the Brunello CRISPRko sgRNA library as pooled library...with both A375 (melanoma) and AT29 (colon cancer) cells and assessed the libraries performance via the depletion... viability screens. Based on these metrics the Brunello library was more effective than other popular ...genes. In the subsampling analysis of the sgRNAs, Brunello even with only a single sgRNA outperformed another...

  8. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...measure, and control molecular signals, cells, and groups of cells in order to understand their activity...Fluorescence-activated cell sorting (FACS): This is a type of flow cytometry that separates mixtures of cells into distinct...Thus, FACS can be used to separate cells expressing GFP from cells that are not. Developmental/transgenic...(GFP) has likely existed in creatures such as jellyfish for millions of years; however, it took until ... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing...consisting of 238 amino acids derived from the crystal jellyfish Aequorea victoria. It has a fluorescent emission... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...

  9. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...of interest into cells. Unlike gamma-retroviruses that can only infect dividing cells, lentiviruses can...dividing and non-diving cells while retroviruses can only infect dividing cells. When choosing the appropriate...to ensure that the proper cell line is used; several common laboratory cell lines including 293 were immortalized...can infect dividing and non-dividing cells.  Addgene has an extensive collection of lentiviral plasmids...transfecting four different plasmids into the producer cells (two packaging plasmids, an envelope plasmid, and...pol, and env), the isoforms of these proteins, as well as the viral long terminal repeats (LTRs) are different...appropriate gene delivery system, the specific target cell or tissue needs to be considered. Q2: Can I use...

  10. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 3:42 p.m.
    ...lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic material...particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor and sometimes ...transduction efficiency, and is an excellent choice for targeting cardiac cells (Bish et al., 2008). It can ... Therapy: An Overview. Cells, 12(5), 785. https://doi.org/10.3390/cells12050785 Kaludov, N., Brown, K....delivery across species. Cell, 184(19), 4919-4938.e22. https://doi.org/10.1016/j.cell.2021.08.028 Tervo, D...tissue tropism, allowing them to target particular cell types. Figure 1: Diagram of AAV components...multiple coreceptors, which may be present on different cell types and sometimes vary between target species ...

  11. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...successfully infected/expressing cells by flow cytometry. The isolated cells were injected into mice and imaged...including monitoring multiple cellular events such as the dynamics of subcellular structures and gene expression...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors...acceptor. NLuc is an excellent BRET donor due to its bright luminescence. Intracellular Calcium Sensor Yang...to changes in intracellular calcium and is useful for direct imaging of events in cells and brain explants...could “clone by phone”. One of my professors was excellent at phone cloning with labs around the world and...luciferase by Promega’s Advanced Technologies group as well as evolution of the coelenterzine substrate to the...

  12. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ... origin as well as a selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both...bee gut microbiome species such as Snodgrassella alvi and Gilliamella apicoli. The plasmids could also ...strategy for bacteria to count cell divisions. The technique, called distributed cell division counting (DCDC...one bright particle that segregates into one cell during cell division. To achieve the agglomeration of ...promoter. Cells are first induced with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are... new particles. With subsequent cell divisions, the fraction of cells containing the particle exponentially...plasmids to both Gram-positive and Gram-negative cells.  The Wang lab further developed a modular suite...

  13. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    Published
    June 15, 2017, 2:30 p.m.
    ...homodimerize in cells. To understand how well fluorescent proteins approach this ideal situation, cellular assays... quantified for individual cells, and the number of 'normal' looking cells can be counted. This assay ...living cells. Strikingly, the OSER assay reveals that tagRFP does not act as a true monomer in cells (Constantini...a non-aggregating fluorescent protein. In the jellyfish Aequorea victoria, AvGFP forms a homodimer. In...tolerated in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic... affinity and has been used to demonstrate that yellow fluorescent protein homodimerization is characterized...fluorescent proteins interact or oligomerize in cells? This question is largely unanswered. Ideally, fluorescent...

  14. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ... Oct 7, 2020 by Gabrielle Clouse. This post was contributed by guest bloggers Marcelle Tuttle and Alex...differentiate induced pluripotent stem cells (Chavez et al., 2015) into neuronal cells by activating genes that cause...drug targets by finding genes that cause cell death in cancer cell lines upon activation (Gilbert et al.,...reasonably well.  Best of luck on your experiments!  Many thanks to our guest bloggers Marcelle Tuttle ...Repression and Activation. Cell 159:647–661 . https://doi.org/10.1016/j.cell.2014.09.029 Konermann S, Brigham...and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/10.1016/j.cell.2014.09.039 Wong N, Liu W,...activation allows for exploratory-based research as well. For example, CRISPRa can be used to find potential...

  15. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...active wavelengths to reach zooxanthellae, the coral’s symbiotic dinoflagellates that are photosynthetic ... them into your cells, you can simply look for blue colonies to determine which cells have taken up the... that the cellular fitness cost varied greatly between chromoproteins. In some cases, cells containing... mScarlet as the donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you... is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful corals...described an unusual blue-colored protein from the jellyfish Cassiopea xamachana (Blanquet and Phelan, 1987...cloning experiment worked just by looking at the cells on the plate. For example, if your plasmid contains...

  16. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...precipitation. Instead of sonicating the cell pellet, I lyse the cells with this AAV lysis buffer and freeze...packaging AAV is transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids...preparing reagents, I choose to only harvest the cell pellet. My yield would be higher if I used PEG precipitation... my experiments, I get enough virus from the cell pellet alone. If yield is important, or if the viral...bath to release the virus particles from the cells. Cell lysates are then treated with benzonase immediately... genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses...five days after transfection, the AAV-containing cells and media are harvested and purified. In total, ...

  17. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...that cells are labeled with their own unique color; and 2) low levels of labeling so that a cell’s projections...tables 1 and 2, with a cell-type specific promoter or enhancer now allows for cell-type-restricted expression...this plasmid pool are injected into an animal with cell-type-specific Cre, the polyA sequence is inverted... library to infect an animal expressing Cre in a cell type of interest, a scientist can isolate DNA from...(s) of the capsid gene successfully infected the cell type of interest. Repeat rounds of infection and...capsid variants with increased ability to infect the cell type of interest, which, in the case of Chan et ...it was actually engineered to target the CNS. How well do these AAVs transduce the CNS and PNS? Both plasmids...

  18. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...efficient gene deletion in B cells, T cells, macrophages, and dendritic cells, with no deleterious effects...to HEK293FT and HeLa cell lines. When the miRNA activity profile for a given cell type is already known...or ZFN to edit immune cells poses challenges such as the inhibition of immune cell differentiation. Furthermore...immune cell activity. However, a CRISPR-based tool for in vivo functional analysis of immune cells has been...construct in bone marrow stem cells from Cas9-expressing mice, and used the cells to reconstitute the immune...vivo using ultrasound in Escherichia coli and Salmonella Typhimurium. This work demonstrates engineered...post-transcriptional gene expression across the majority of cellular pathways. Despite several years of research, the...

  19. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...specific cell types (populations) within neuronal networks in living animals. To accomplish such cell specific...individual cells. With this technique, a Neuroscientist can see the full dendritic arbor of a single cell with...Cre-mediated possibilities for site specific (and often cell type specific) control of DNA recombination and ...expression can be advanced by the coordinated use of fellow SSR system FLP-FRT. In addition, a variety of means...30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular evolution...revolutionize neuroscience Manipulation of specific cell types in neuronal networks Double inverted vectors...combination with mouse lines that express Cre only in the cell type of interest (Atasoy et al. 2008; Cardin et ...

  20. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...fluorescence activated cell sorting (FACS) to isolate differentiated from undifferentiated cells at the end of ...these cell types are used. The lack of good flow cytometric antibodies for many CNS-resident cell types...for which CRISPRa is well-suited, including studying mRNA dosing effects, cellular reprogramming, and disease...that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed PMID: 30318302...-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center. During...relationships in defined substructures of the CNS as well as individual neuronal populations.  The Yang Lab...lentiviral vectors for in vitro and in vivo CRISPRa (Savell et al., 2019). This dual lentivirus system pairs...
Showing: 141 - 160 of 845 results