We narrowed to 818 results for: cell
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TypeBlog Post...percentage of total live cells are GFP+. In this plot, with adjusted voltage, cells that did not receive ...would serve you well. Percentage of cells in a parent gate (all live cells for example) that are positive ...example, you have introduced a GFP protein into your cells (visualized via the FITC channel) and you are trying...graphs could simply be the percentage of the total cells present in the gate. Figure 1: Example...looking at proteins that are expressed regularly in cells and might be upregulated or downregulated based ...populations. MFI is the average brightness from all cells that are positive for the marker of interest. MFI...
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Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS
TypeBlog Post... to express one gene in one cell type and another gene in a different cell type (6–8). Although it’s often...systematically study gene function with temporal control and cell-type specificity! The Gal4/UAS system At a more...the hedgehog gene can be overexpressed in several cell types by using different Gal4 drivers. It is much...much easier to mate flies to get the right cell type expression as opposed to making a new fly with hedgehog... Targeted gene expression as a means of altering cell fates and generating dominant phenotypes., Development...Neuro-Trauma during Asymptomatic Viral Infections, Cell Host Microbe 22, 48-60 (2017). PubMed PMID: 28704652...expression, lineage tracing, and mosaic analysis., Cell 141, 536–48 (2010). PubMed PMID: 20434990. PubMed... -
Plasmids 101: Stringent Regulation of Replication
TypeBlog Post...replicating too quickly can slow cell growth or even kill the host cell. For stringently controlled plasmids...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...be synchronized with the replication of the host cell's chromosomal DNA. Plasmids are said to be under... stringent control when they depend on the host cell’s initiation proteins to start their own replication... occurs at the appropriate time—for example, the cell must have enough nutrients available to complete...the amount of active DnaA available is one of a cell’s best tools to regulate DNA replication. DnaA negatively...energy source, ATP binding is one more way for the cell to control DNA replication. A variety of other proteins... -
Transferable Skills Guide: Time Management
TypeBlog Post...splitting my cells first. Time management through planning I probably wouldn’t have fallen into my cell splitting...for Scientists. Back when I used to do mammalian cell culture, I would often respond to friends asking...there in a few minutes, I just need to split some cells.” Inevitably, this task would take longer than I...since my days as an undergrad working with cancer cell lines. As I started planning out my own experiments.... In my anecdote above, I prioritized splitting cells over meeting up with friends, but didn’t realize...mean in terms of the utility of your tasks. In my cell splitting anecdote, I probably could have grabbed...with friends and come back to the lab to split my cells later. Eating with friends has a somewhat immediate... -
mTOR Pathway
TypeCollection...Laplante M, Sabatini DM. Cell. 2012 Apr 13;149(2):274-93. doi: 10.1016/j.cell.2012.03.017. PubMed PMID... (mTOR) is a key metabolic regulator controlling cell growth and proliferation. mTOR, a serine/threonine...signals to determine the nutritional state of the cell and regulates the switch from anabolic to catabolic...growth factors, including insulin. mTORC2 influences cell survival and cytoskeletal organization. Cancer Impact... promotes the conditions needed for uncontrolled cell growth and proliferation, including an abundant ...appears to signal upstream of Akt, also promotes cell survival and proliferation. mTOR Pathway Plasmids... one drug, many effects. Li J, Kim SG, Blenis J. Cell Metab. 2014 Mar 4;19(3):373-9. doi: 10.1016/j.cmet... -
Promega Plasmid Collection
TypeCollection...expression systems including E. coli , mammalian cells, and cell-free systems. HiBiT Fusions HiBiT is a small...proteins at endogenous levels in lysates or live-cells, thus preserving physiological relevance. NanoBiT... to either LgBiT or SmBiT and expressed in live cells. When the two proteins interact, the subunits come...designed to measure molecular proximity in living cells. NanoBRET PPI assays use NanoLuc Luciferase as a...NanoBRET 618 fluorophore, as the energy acceptor. Live-cell reagents allow you to detect PPI induction and inhibition... NanoLuc luciferase-fusion protein expressed in cells. Additional Resources Browse Addgene's Luciferase...NanoLuc fusions are useful for both intracellular and extracellular applications where you want minimal... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog Post...when trying to screen a limited number of cells like primary cells or in vivo. Recently, the labs of David... cultured cells as well as more difficult conditions such as model systems and primary cells. References...with both A375 (melanoma) and AT29 (colon cancer) cells and assessed the libraries performance via the depletion...important criterion for probing a limited number of cells. Get the Brunello CRISPRko sgRNA library as pooled...subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition (CRISPRi) libraries...study screening for vemurafenib resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation...CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. PubMed PMID: 25307932. PubMed... -
AAVs in Retinal Gene Therapy
TypeBlog Post...expression in non-dividing cells because they remain in episomal form in the cell’s nucleus. But since nothing...hope are many more clinical successes for gene (and cell) therapies. One may wonder why the first AAV-based... providing long-term gene expression in retinal cells - AAV - were becoming available. At the same time... lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of choice for ocular...long term expression in dividing and non-dividing cells due to its (random) integration into the genome,...limited tropism in the retina, primarily infecting cells of the retinal pigmented epithelium (RPE). AAVs... for patients, this gene can be targeted to RPE cells using a lentiviral vector and a clinical trial is... -
Plasmids 101: Multicistronic Vectors
TypeBlog Post...them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially if you...reporters allow you to easily screen or select for cells that are expressing your gene of interest at high...promoter, multicistronic plasmids ensure that any cells that are positive for your marker should also be...anytime you want to express multiple genes in the same cell. Although it is possible to drive co-expression ...promotes translation of the viral genome in eukaryotic cells.1,2 Since then, a variety of IRES sequences have... cistron due to factors such as the experimental cell type and the specific genes cloned into the vector...scientists. 2A cleavage is universal in eukaryotic cells, and, although some scientists report close to 100%... -
An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success
TypeBlog Post...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed ...Cepko Interested in Working on the Eye 9:05-10:16 : Cell Targeting in Retinal Gene Therapies 10:17-14:34 ...28:39 : Methods for Delivering Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools...publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation with GFP-dependent Cre...