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Showing: 191 - 200 of 872 results
  1. Quick Guide to All Things Lentivirus

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    ...-dividing cells. The ability to infect non-dividing cells is not restricted to in vitro cell culture as...target cells. If you are transducing non-adherent cells you might need to centrifuge your cells with the...favourite gene into your favourite cell line or into primary cells, this blog article will give you some...infect both dividing and non-dividing cells, including stem cells. These properties make lentiviral vectors...immortalized cells lines but reach their limits when it comes to transferring DNA into primary cells or non-...non-dividing cells. To target these un-transfectable cells, labs use viral vectors and particularly retroviral...will first need to transfect producing cells, usually 293T cells, with your transfer plasmid and your packaging...
  2. Plasmids 101: Protein Expression

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    ...produced using complex mixtures derived from cells or using live cells. There are several types of expression...consists of transfecting cells with your DNA template of choice and allowing these cells to transcribe, translate...demanding cell culture conditions compared to other systems. Insect expression systems Insect cells can also...coli or yeast cells. The only problem with baculovirus systems is that the infected host cell is eventually...eventually lysed. Cell lysis halts protein production, but there are non-lytic insect cell expression systems...Sf21, Hi-5 cells)  that allow for continuous expression of genes integrated into the insect cell genome. ...that is then transformed into a bacterial cell. Transformed cells propagate, are induced to produce your ...
  3. Getting the Most from Your Lentiviral Transduction

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    ...Love your cells and they'll love you back A successful lentiviral infection begins with your cells, both ...both your packaging cells and your target cells. Poor tissue culture practices can lead to low titers and... promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...reaching 108 cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients... your producer cells with fresh medium. Daily medium exchanges help alleviate cellular stresses associated...consumption due to the high cell density of the culture. In addition to benefitting your cells, medium exchanges...
  4. What the HEK?

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    ...embryonic kidney (HEK) cells originated from? Or why they are such a commonly used cell culture model? In ...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first ...inconsistent with kidney cells (Shaw et al.). Since then, transcriptome studies of the cell line have determined...for?   As one of the older cell lines in the research community, many cellular research tools have been ...any unwanted material (HEK cells, etc.) and are ready to use on another cell or organism. This pipeline...where HEK cells may be particularly useful due to their intrinsic properties. HEK-293 cells are tumorigenic...researchers using the cell line. What the HEK is wrong with it?   All cultured human cell lines are susceptible...
  5. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    ...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...white inclusions in cells with GVs via phase contrast microscopy, GVs visible in cells via TEM, and purified... heterogeneity required single cell screening to isolate a clonal cell line capable of producing ultrasound...expression homogeneity, as the cells could be batch-sorted to isolate a polyclonal cell line with strong gas vesicle...transient transfection in rtTA+ cell lines (such as HEK293-TETOn cells), or they can also be integrated...PiggyBac transposase. In rtTA-cell lines, it is better to first engineer the cell line with rtTA, then integrate...own mARGAna cell line. (c) Z-projected phase contrast images of GV expressing MDA-MB-231 cells (top, scalebar...
  6. Hot Plasmids - October 2020

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    ...determine cell cycle length by examining the ratio of blue to red cells. Cells undergoing rapid cell cycles...engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. They used...’s difficult to observe the cell cycle in vivo, particularly if the cells are migratory or in deep tissues... BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations of IL-3. Cells grown...proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in media without IL-3 show...H2B-FT consistently predicted the cell cycle speed across these different cell types. In one example, H2B-FT...expressed in murine B cells (BaF3), which depend on IL-3 to proliferate. The cells cultured with a low ...
  7. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    ...infected cells, the other using wild-type Cas9 to remove the HIV-1 genome from infected cells. Read on...1 reservoirs HIV-1 infects cells in the immune system, notably CD4+ T-cells, and eventually leads to acquired...percentage of cells with activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that... genome from every infected cell and 2. prevent reinfection of those cells. Kaminski et al. designed gRNAs...Kaminski et al. observed no negative effects on cell viability, cell cycle progression, or apoptosis with Cas9...in HIV-1 infected T-cells to see if they could be rescued by CRISPR. CD4+ T-cells were isolated from healthy...1 is limited to a subset of T-cells, but it is unclear if enough T-cells could be reached to ablate a ...
  8. Antibodies 101: Introduction to Antibodies

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    ...on or in these cells: antibodies can bind to proteins on the cell surface, or the cell membrane can be...antibodies are produced by B cells. They bind to proteins on the surface of extracellular pathogens such as parasites...fusing B cells that have been extracted from animals injected with protein antigen to myeloma cells. The ... measurement of protein expression in cell lysates, whole cells, or tissue samples. Many experiments that...heterogeneous mixture - for example, cell lysate or media from cultured cells. ELISA can be used to quantitatively...same cell simultaneously as well as compare between samples. Learn more about flow cytometry. Cell and ... (immunohistochemistry) or cells (immunocytochemistry). This could be cells or tissue slices mounted to...
  9. 27 Hot Plasmids from 2016

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    ... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ...plasmid to generate a population of mutant cells. The mutant cells were treated with WNV at a dose and duration...regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3, EMC2, DERL2, UBE2G2...small GTPases involved in cell signaling pathways that control many cellular processes such as differentiation...therefore varies with cellular location: it is both red and green through much of the cell, but turns more ...AID-tagging system to essential genes in human cells and mouse ES cells because it was difficult to fuse the AID...-to-study genes in hard-to-study cell types. Natsume T, et al. Cell Rep. 2016. PubMed PMID: 27052166 ...
  10. Viral Vectors 101: Pseudotyping

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    ...infecting most cells, but not all. For example, LDLR expression in hematopoietic stem cells, T cells, and B cells... host cell. Some of these envelope-receptor pairings are broad, allowing delivery into many cell types...delivery to only a few cell types.  If you’re looking for more control in the cell types your lentiviral...protein causes cells that it infects to fuse together and form syncytias that cause the cells to die. Cytotoxicity...Cytotoxicity can also be cell type dependent: if a cell has few receptors for a specific envelope protein...lipoprotein receptors (LDLRs) on a cell surface to endocytose into the cell. The reason VSV-G is so commonly... cells is low. Infection of these cells with VSV-G pseudotyped virus requires the upregulation of LDLR...
Showing: 191 - 200 of 872 results