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We narrowed to 813 results for: cell;

Showing: 121 - 140 of 813 results

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

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    Blog Post
    ...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..., these reporters can help define the different cells present in a mixed population, or identify differentiation...turning the single-input reporters into multi-input cell classifiers in one assembly step. Find the miRNA...

  2. Viral Vectors 101: Types of viruses

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    Blog Post
    ...dividing cells only, so is not suitable for infection of quiescent cells or infrequently cycling cells but ...non-dividing cells, making it conducive to targeting all cell types. Altogether, AAV has many excellent features...DNA copies of their genome upon infecting a host cell. This viral DNA (also called proviral DNA) is then...and permanently integrate a genetic cargo into a cell. These viruses have a cargo capacity of ~ 8 kb, ...needs, and can transduce (infect) a wide range of cell types with a high efficiency. However, gamma-retrovirus...but is fine for experiments with immortalized cell line cultures. Lentivirus Lentivirus is a type of retrovirus...lentivirus can infect both dividing and non-dividing cells. Lenti particles enter the nucleus through nuclear...

  3. Viral Vectors 101: Preparing Pooled Libraries

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    ...transduction of cells, resulting in cell population mutated by the viral pooled library. These cells can then...transduce your cells.  Titering After generating your lentivirus, you’ll want to infect your cells with the ...of a cell being infected by only one viral particle, but it also increases the chances that a cell doesn...adequately represented in the cell population, you’ll need to infect many more cells than the number of plasmids...appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus. Purification of...small aliquot is not enough to transfect producer cells, so your first step will be to amplify the library...approach for producing lentivirus in Lenti-X 293T cells using a polyethyenimine transfection protocol. I...

  4. Antibodies 101: Producing Recombinant Antibodies

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    ...and yeast cells are cost-effective and faster-growing than mammalian cells, but mammalian cells can perform...your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture...production system, cell lines typically used include Human Embryonic Kidney (HEK) cells and Chinese Hamster...recombinant antibodies with some molecular biology and cell culture experience. Let’s go over the basics of ... place in bacterial, yeast, plant, or mammalian cells. Each of these systems has their advantages and ...post-translational modifications that bacterial and most yeast cells cannot. Your choice of production system may depend...Hamster Ovary (CHO) cells. (L'Abbé et al., 2018).  Recombinant antibodies can be produced through a single...

  5. CRISPR History and Development for Genome Engineering

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    ...genomic loci in living human cells by an optimized CRISPR/Cas system. Cell . 155(7):1479-91. PMID: 24360272... for Mapping Multiprotein Complexes in Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander...described CRISPR-based genome editing in mammalian cell culture. Five years later, PubMed lists more than...multiple loci can be easily expressed in the same cell or organism. The ability to precisely target CRISPR...genome edits or double-strand breaks. In mammalian cells, Cas13-ADAR2 does not require a specific flanking...proteins can be used to image genomic loci in live cells. RNA targeting : Cas13 enzymes target RNA rather...non-specific cleavage does not occur in mammalian cells. Purify : Epitope-tagged dCas9 can also be used ...

  6. CRISPR in the Clinic

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    ...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (2013). ...patient’s own cells that have been modified ex vivo, previously edited healthy donor cells can be delivered...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (2013). Gerald Schwank et al. ...on editing animal models of disease and cultured cells. For example, in 2013, Jinsong Li and colleagues...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis ...delivery directly to the patient. Patient-derived cells can alternatively be harvested, edited ex vivo, ...therapies involve editing healthy donor-derived cells and delivering them to the patient.   Current...

  7. Neurodegeneration Research Collection

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    ...protein-protein interactions and nerve cell function within the cell. Parkinson's Disease Parkinson’s disease...inducible vector . Harley et al. Cell Rep. 2023 Dec 26. Induce CNS cells with concomitant expression of ...that causes the progressive degeneration of nerve cells in the brain. Symptoms include changes in behavior...FRET-based reporter to study tau seeding in mammalian cells. Lathuiliere et al. Mol Neurodegener. 2023 Aug 8...system, peripheral nervous system, vesicular brain cells and more. New and Noteworthy: AAV.rTH.PI.Cre.SV40...and cortical neuron differentiation. Held et al. Cell Rep. 2023 Aug 30. Enhance differentiation to mesencephalic...neuronal identification in C. elegans. Yemini et al. Cell. 2021 Jan 7. Use a FRET-based method to monitor ...

  8. Plasmids 101: Plasmid Incompatibility

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    Blog Post
    ...copy number in the cell is high, but allows replication when copy number in the cell falls too low (Novcik...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...eukaryotic cells). The partition NTPase is recruited and “walks” each plasmid to a separate daughter cell. When... daughter cell, instead of partitioning each type of plasmid into a separate daughter cells (Figure 2)...from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...different plasmids to be maintained in one bacterial cell. In this Plasmids 101 post, we’ll cover why this...incompatibility occurs when multiple plasmids within one cell have the same replicon and/or partitioning system...

  9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

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    ... and zap the mixture of DNA into cells. Unfortunately, not every cell successfully takes up plasmid DNA...selection: Cells that have gained a specific gene survive In positive selection, only cells that contain...cassette, you would plate the cells on kanamycin-containing agar. Only cells carrying plasmids that contain...Therefore, your cloning strategy needs to identify cells containing the plasmid construct you’re seeking....resistance marker. After transformation, plate the cells on medium containing the corresponding antibiotic...these cases, antibiotic selection will select for cells that have been transformed with the plasmid backbone...plasmids to complement auxotrophy. Negative selection: Cells that have lost a specific gene survive Unlike positive...

  10. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    ...hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells can be implanted...Annotation of genes in immune cells typically involves the creation of germline knockout mice, which ...CRISPR-based systems enable gene knockout in immune cells in a high-throughput manner, but these systems have..., ubiquitous knockout of single genes in immune cells in vivo (LaFleur et al., 2019a and LaFleur et al...many genes have different functions in different cell types, at different times, and in combination with... L-CHIME Knocks out a single gene in defined cell types (lineage-based) pXPR_070 Violet-excited...benchmarked on positive control genes in CD4+ and CD8+ T cells. To demonstrate the power of these systems to investigate...

  11. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

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    Blog Post
    ...identify genes essential for cell viability in cancer and pluripotent stem cells. In a melanoma model, they...high representation of library within cells and aimed for 300 cells per construct, but in some biological...were able to produce a very diverse population of cells within a lab dish in a matter of a few days. Their...pooled libraries include enrichment against kinases, cell cycle proteins, and more.  “In the past, genome ... delivered via lentiviral infection into target cells that have previously been stably transfected for...conduct systematic genetic screens in mammalian cells,” David Sabatini, a senior author of one of the ...’s also important that you infect enough target cells. What’s “enough” depends on your experiment and ...

  12. Plasmid Tools for Microbiome Studies

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    Blog Post
    ...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... and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated with ...this system in vitro and in vivo. They identified cells from 19 genera across four phyla that have taken...

  13. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

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    ...introduced the reporter into the cells, you’re ready to edit. These cells are then transfected with several... up, all successfully transfected cells will express RFP while cells containing the reporter gene that...using the RFP:GFP ratio, edited cells can be sorted from unedited cells. In an experiment that targeted...Activity. Frontiers in Cell and Developmental Biology 6: . https://doi.org/10.3389/fcell.2018.00054 Sander...CRISPR experiment was successful in your organism or cell type? You can use DNA sequencing or other molecular...histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct...simply changing the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The Madison lab ...

  14. Lentiviral Vector Uses and Overview

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    Blog Post
    ... of cell types, so they’re ideal for work with primary cells or other difficult-to-transfect cells. Lentiviral...dividing cells. Another type of vector, based on adenovirus, could infect non-dividing cells, but without...ability to transduce both dividing and non-dividing cells. Lentiviral vectors are the vector of choice for...lentivirus HIV-1, well-known to infect non-dividing cells. The first lentiviral vector system consisted of...viral envelope to dictate the tropism, or types of cells the vector could infect. Finally, the transfer plasmid.... After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles...adeno-associated viral vectors can also target non-dividing cells, lentiviral vectors can carry much more genetic ...

  15. Ras Pathway

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    ...molecular switches that transmit signals within the cell. GTPases have an active (GTP) and inactive (GDP)...prominent roles in signaling pathways regulating cell migration, survival, adhesion, growth, and differentiation...Cancer Impact Ras is ubiquitously expressed in all cells and all tissues. Ras can activate a variety of ...pathways, which ultimately turn on genes involved in cell growth, differentiation and survival. Ras is essential...essential for regulating the growth of the cell, however when mutations occur in the Ras gene it can be... active form, constantly activating pathways for cell growth and potentially leading to cancer. Constitutively...E2F2 E2F3 E2F transcription factor ECT2 Epithelial cell transforming 2 EGFR Epidermal growth factor receptor...

  16. Fluorescent Protein Guide: Subcellular Localization

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    ...between animal cells. They are important for holding cell structures together and reducing cell stress. There...give the cell its shape, organize cellular organelles, and are necessary for movement and cell division...example, houses the cell’s genetic material. The mitochondria produces the energy cells required for chemical... on cell cycle) Centrin-2 EGFP Erich Nigg 41151 pEGFP Cep170 C-term Centrioles (dependent on cell cycle...Fluorescent Protein Collection . Return to top Cell Junctions Cell junctions are responsible for forming connections... General Structures The cell contains a variety of organelles and subcellular structures each with their...its role in secreting proteins from the cell to the extracellular environment. Proteins are first translated...

  17. Antibodies 101: Monoclonal Antibodies

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    ...isolating the B cells from an immunized animal, those B cells are fused to myeloma cells at a 1:1 ratio...fusion between an antibody-producing B cell and a myeloma cell (National Research Council (US) Committee...immortal cell population that will continue to produce the specific antibody that the original B cell produced... selects against unfused myeloma cells and antibody-producing B cells that cannot replicate, meaning that...monoclonal antibodies are derived from a single B cell clone. One common way of producing monoclonal antibodies...with BioRender.com.   The antibody-producing B cells that are used to make hybridomas come from animals...antibody-encoding genes over time as the hybridoma cells divide. These changes can alter the antibody that...

  18. Predicting Adverse Reactions to Monoclonal Antibody Drugs

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    Blog Post
    ...epitopes likely to be presented by T cells to elicit an immune response. T cell epitopes are found in either ...that take place within an antigen-presenting cell like a T cell. It is also not yet possible for these algorithms...traditional 2D assays with immune cell lines or peripheral blood mononuclear cells (PBMC) in 3D models of the...CD4+ or CD8+ T-cells or monocytes. PBMCs can be further differentiated into dentritic cells or macrophages...impurities in the formulation. Efforts to predict T cell epitopes focus on characterizing those epitopes ..., that are commonly used for the prediction of T cell epitopes based on the amino acid sequence of the...lymphatic system. PBMCs consist of various immune cells, and can be used as a whole or can be used to isolate...

  19. Custom CRISPR Screens & the Green Listed Software

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    ... the single cell level), are separated by, for example, cell sorting. The separated cells are subsequently...more than 8x107 successfully targeted cells. Depending on the cells used and the readout, this could be ...My group works on immune cells in different contexts. Most of the immune cells we study are migratory, ...CRISPR modified cells are transferred into an animal and the behavior of the transferred cells is followed...genes essential for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683. PubMed PMID:...with an easily measureable cellular phenotype (e.g. survival of a cancer cell as a cytotoxic drug is added...With these screens, the investigator generates a cell population where all genes are knocked out at the...

  20. Adenoviral Vector Production and Troubleshooting

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    ...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity...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...stock by doing a plaque formation assay on A549 cells . Alternatively, antibody-based assays are commercially...
Showing: 121 - 140 of 813 results