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We narrowed to 877 results for: cel

Showing: 341 - 360 of 877 results
  1. Google Forums Round Up: First Impressions of NgAgo

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    Blog Post
    ...degraded by the cellular machinery, leaving a small amount of active NgAgo/ssDNA complex in the cell. As proposed...results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL NgAgo ARTICLE... guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA ...dangerous, non-physiological temperature for mammalian cells and one which diminishes the endonuclease activity...guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid.... Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or...lipofectamine-based method to deliver NgAgo ingredients into the cell, the same method used by Gao et al, 2016 (1). View...
  2. AAVs in Retinal Gene Therapy

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    Blog 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... gene restores vision in the dog model of LCA (Lancelot, one of the dogs who received the AAV-RPE65 treatment... 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...
  3. MXS Chaining

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    Blog Post
    ...protein localization, as well as cellular processes like gene expression and cell migration (Sladitschek and... MXS-chaining were used to label subcellular structures in live cells. Four individual expression constructs...construct was then introduced to HeLa cells (Figure 1). The resulting HeLa cells show robust labeling, with strong...imaging or flow cytometry applications in mammalian cells. The modules used in this method include fluorescent...components, one can generate constructs to visualize cell cycle dynamics, titrate inducible transgene expression...detection of each fluorophore at the respective subcellular structure (Figure 1). Here, MXS Chaining gave...kb insert. Table 1: MXS Constructs for Labeling Cellular Structures Construct Fluorophore Excitation...
  4. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

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    Blog Post
    ...Stability improvement of antibodies for extracellular and intracellular applications: CDR grafting to stable...systems, and often do not fold properly once inside cells.  Developing the HA frankenbody To combat the issues... detects the widely used HA epitope in diverse cellular environments. They called this probe the HA frankenbody...previously proven to fold and function in living cells. To make these scaffolds specific to different targets...the tagged protein upon probe binding in living cells.  In their proof of concept experiment the Stasevich...mCherry, 4xHA tagged histone protein (H2B) in U2OS cells. They then showed that the HA frankenbody probe ...that the HA frankenbody is effective in sensitive cells such as living neurons, and can be fused to other...
  5. Viral Vectors 101: What is a Virus (and a Viral Vector)?

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    Blog Post
    ...hijacking the machinery of the cells they infect. Once they enter a cell, they treat their viral genome...genome as a “payload” that is delivered to the cell. The cell then uses its own machinery to replicate the...replication process whereby one cell copies itself and “splits” into two cells. Viral vectors Viruses are ...virus. From one, come many After a virion infects a cell, the single particle creates many copies of itself...millions of new copies, which are released from the cell like baby spiders from an egg sac. (Okay, some enveloped...lysing, but they do generally still kill the infected cell.) This is in contrast to bacteria and eukaryotes...means the vector can deliver a genetic payload to a cell but cannot create new particles. As a bonus, this...
  6. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    ...relied on for transducing cells: galactose and the AAV receptor (AAVR). Cell lines lacking galactose were...Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6 (2007): 1339-1347... Wilson labs determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B...commonly used to enrich adult mouse hematopoietic stem cells. Its ligand(s) has not been identified. While LY6A...out of Ly6a in primary C57BL/6J brain endothelial cells decreased transduction rates while overexpression...overexpression of the C57BL/6J version of Ly6a in 293T cells enhanced PHP.B transduction rates. The group also determined...PHP.B, virus still reached the brain endothelial cells of these mice, although at lower levels than in ...
  7. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...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...that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed PMID: 30318302...CRISPRa screening approach in embryonic stem (ES) cells to determine which transcription factors promote...antibodies, downstream of the TUJ1 coding sequence. ES cells that stably express the dCas9-Suntag activator were...two weeks to allow differentiation to occur. CD8+ cells were then isolated and their genomic DNA sequenced...also be applied to the differentiation of other CNS cell types, including microglia, astrocytes, and oligodendrocytes...
  8. CRISPR Plasmids - gRNAs

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    Collection
    ...is highly recommended that you determine if your cell type contains the target sequence before using any...
  9. CRISPR Plasmids - Cascade-Cas3

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    ...bound to DNA, and the break can be repaired by the cell’s endogenous repair mechanisms. Cas3 belongs to the...
  10. Brzezinski Lab CRISPR Collection

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    ...enhancers in the retina alters photoreceptor and bipolar cell fate specification, yet differs from deleting both...
  11. Using CRISPR/Cas9 to Edit Disease Out of the Genome

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    ...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (5 December...CRISPR in adult stem cells has huge potential, because the genomes of adult stem cells in organoid cultures...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (5 December 2013). 8. Gerald Schwank...homologous recombination in cultured intestinal stem cells of patients with cystic fibrosis (8).   The difference...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis....is a big advantage over induced pluripotent stem cells, which need to go through a phase of crisis where...B. 2021. “CRISPR/Cas gene therapy.” Journal of cellular physiology.  https://doi.org/10.1002/jcp.30064...
  12. Hot Plasmids - February 2022

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    Blog Post
    ... including live-cell imaging, single-molecule tracking, immunolabeling of fixed cells, Western blotting...possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular Cell. 2021. https://doi.org...Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized genetic screening technique ...Anti-FLAG Frankenbodies for modular imaging of cellular targets by: Mike Lacy Adding to their set of anti-HA...well-folded scFv scaffold to improve its stability in cells, and use it in a variety of applications. This set...frankenbodies to amplify the signal.  Liu et al. Genes Cells. 2021. https://doi.org/10.1111/gtc.12893, Zhao et...Cas12a, creates challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature...
  13. A Practical Approach to Choosing the B(right)est Fluorescent Protein

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    Blog Post
    ...dodge cellular variation observed in mammalian cells is to tag an endogenous gene. By imaging cells (or ...mammalian cells within the dish. This is caused by the stochastic nature of plasmid uptake by cells. To correct...intended application. The three key properties for live cell imaging that will be treated in this series of posts...Determining practical brightness When a dish of mammalian cells is transiently transfected with a plasmid containing...allows for a precise determination of brightness in cells, under realistic experimental conditions. By determining...al (2017) and Heppert et al (2016). In mammalian cells this would be possible with CRISPR/Cas-based genome...efficiency of the fluorescent protein in the host cell, which is a key determinant of the practical brightness...
  14. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    ...human T cells and hematopoietic stem and progenitor cells (HSPC)s to ~40% expression in the K562 cell line...gene to airway epithelial cells in mice. This vector transduced ~10% of cells, similar to the transduction...Using AAV Donor Vectors. Cell Reports 20:750–756 . https://doi.org/10.1016/j.celrep.2017.06.064  Carvalho... then independently packaged into an AAV. When a cell is co-infected with both part A and part B AAVs,... always.  The combination of serotype and tissue/cell type could have a big impact on the rate of transgene...since both of these variables influence whether cells will be co-transduced with multiple copies of both... followed by the 3’-end of the transgene. When a cell is transduced by both viruses, if the AAV genomes...
  15. Treating Muscular Dystrophy with CRISPR Gene Editing

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    ...persist over time, stem cell editing would be highly desirable. Should stem cell editing occur, the CRISPR... high percentage of muscle cells throughout the body, especially stem cells 2. Remove any immunogenicity...patient myoblasts; when injected into DMD mice, these cells expressed functional dystrophin. Editing genes in...in vivo is of course much more difficult than in cell culture, but Ran et al. have shown that CRISPR and...be delivered to both cardiac and skeletal muscle cells, where precision editing of the dystrophin gene ... the percentage of dystrophin-expressing muscle cells increases over time, and all three groups confirm...editing. Tabebordbar et al. show that muscle stem cells are modified by this approach, increasing the possibility...
  16. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using the U-containing strand..., which may lead to cell death rather than NHEJ repair, thus removing these cells from the edited population...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual base...relatively low frequency, especially in nondividing cells. Scientists everywhere wistfully imagined being ... and DNA repair. Black arrows indicate natural cellular processes. BER = base excision repair. Note: hypoxanthine...editing efficiency, Komor and team wanted to force the cell to use the deaminated DNA strand as a template. ...frequency to above 30% for a variety of targets in human cell lines, with an average indel frequency of only 1.1%...
  17. Choosing the B(right)est Fluorescent Protein: Photostability

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    Blog Post
    ...intensity evolves over time in living cells at a power used for live-cell imaging. To conclude, the most photostable...molecule will photobleach at some point. For live-cell imaging, it is desirable to have fluorescent proteins...experiment. The best fluorescent proteins for live cell imaging can be excited many times, thereby producing...these factors, environmental conditions such as cellular redox state and oxygen concentration may affect...protein is a key requirement for quantitative live-cell imaging, it is important to be able to quantify ...it is advisable to perform time-lapse imaging of cells producing the fluorescent protein of interest under...natural situation. Measuring photostability in living cells provides a much more realistic view of photostability...
  18. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    ...genes essential for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683. PubMed PMID:... (and multiple) genetic perturbations in single cells. Methods to scale up multiplexed CRISPR systems ... build and constantly improve gRNA libraries, excellent resources with effective gRNAs are publicly available...perturbations and applications in functional genomics, cell reprogramming, and beyond. For further details, ...CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. PubMed PMID: 25307932. PubMed...Genes and Genotype-Specific Cancer Liabilities." Cell 163.6 (2015): 1515-1526. PubMed PMID: 26627737. ...Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library." Nature...
  19. Resolute Plasmid Collection

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    ...quality of research output, RESOLUTE expects to accelerate the pace of research in the field of SLCs to ...
Showing: 341 - 360 of 877 results