We narrowed to 158 results for: mec 1 cells

...labeling T cells, B cells, and antigen presenting cells, scientists have been able to decypher cellular dynamics...tracked by cell sorter giving immunologists the ability to count cells and statistically analyze cell movements...fast T cells could jump from one antigen presenting cell to another. Indeed, we now know that T cells are...separates the cells mechanically in a vibrating nozzle, imparting a positive or negative charge to cells that ...2016)? How do dendritic cells migrate to lymph nodes to activate T cells (Celli et al 2008, Kitano et al...In complex metazoans, rapid cell division and large scale cell mobility are essential processes during.... a basal progenitor cell migrating to the epithelium). At this stage, only cells from the immune system...

...more applications.   Fig. 1 Making a Protein Tag (Abridged) (1) Locate a promising monoclonal antibody...For about as long as the disciplines have existed, cell and molecular biologists have been continuously ... peer into the mysterious, microscopic world of cells. The development of antibodies as tools allowed ...monoclonals would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different...specific for human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616. https://doi.org/10.1128/mcb.5.12.3610... and immunoglobulin heavy chain binding protein. Cell 46:291–300. https://doi.org/10.1016/0092-8674(86...signal prevents secretion of luminal ER proteins. Cell 48:899–907. https://doi.org/10.1016/0092-8674(87...

...currently doing this using two approaches: single-cell RNA-sequencing to identify neuronal class and deep...WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans embryo when all essential...Wang et al., 2018).” With the split system, only cells that have both halves of cGAL can drive expression...Zhang et al., 2015). In this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin...splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein domains that naturally...mutations with unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust...C.elegans neural network, and Sibernetic, a fluid mechanics simulator that can be used to simulate muscle ...

...kinase essential for cell proliferation signaling. CRISPR knock-in HiBiT-tagged EGFR cells allowed researchers... carried by extracellular vesicles (EVs), edited transcripts could endow neighboring cells with pro-oncogenic...Degradation of Misfolded CFTR. Cells, 12(23), 2741. https://doi.org/10.3390/cells12232741  Kamio, T., Kono, Y.,... fusions utilizing bulky tags can disrupt normal cell biology and make it hard to capture true protein...researchers with a means to quickly iterate, work across cell models, and even introduce mutations. This vector-based...and other fluorescent proteins revolutionized live-cell protein detection, their bulk (~27 kDa) can perturb...Bioluminescence detection is a cornerstone of modern cell-based assays and enables ultra-sensitive detection...

...Making contact with a cell Although an individual AAV particle’s ability to infect a cell is dependent on its...construction and the cell it’s infecting, getting these particles to infect cells is also very titer-dependent... particle to infect a cell, it has to come into physical contact with that cell. Since there is a limited...of surface area on each cell, the way to get more AAV in contact with the cell is to increase the AAV ...between the cell and any individual AAV particle is rare. But simply getting into the cell may not always...particles that have entered the cell now have to travel up the axon, through the cell, and potentially spread ...particles per cell, you may not be able to detect AAV-mediated gene expression in a target cell. Practical...

...the producer cells and purification protocols often describe a process that begins with cell lysis to release... steps to separate cell culture supernatant containing exo-AVVs from producer cells, and then to enrich...effectively target less accessible cells, such as brain tumor stromal cells in a mouse model of glioblastoma...particles is able to infect more cells (as demonstrated in different cellular environments such as retina ...Exosomal vesicles secreted from producer cells also contain cellular macromolecules (e.g. proteins, nucleic...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production... to be spontaneously released from the producer cells into the supernatant during the production phase...

... the cell bodies of the projection neurons reside.   Impact for scientists While the mechanisms by which...they either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...particles require receptor-mediated entry into the cell. This means that not all neurons and pathways will... There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to...significantly updated in 2023 by Jason Nasse.  References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant...

...Inhibitor of CRISPR-Cas9. Cell 170:1224-1233.e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha...Off-Switches for CRISPR-Cas9. Cell 167:1829-1838.e9 . https://doi.org/10.1016/j.cell.2016.11.017 Shin J, Jiang...concern is that the longer Cas remains active in a cell, the greater chances there are for off-target edits...within the first six hours. When added to human cells six hours after the introduction of Cas9 RNPs, AcrIIA4...CRISPR/Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development...cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can block CRISPR activity... ways to anti-CRISPR proteins in your experiments 1. Reduce off-target effects Prolonged Cas activity ...

...development. Single cell sequencing-based lineage tracing Recently, gene expression in all cells of the developing...proportions of otherwise normal cell types are produced, rather than novel, abnormal cell types. Sequencing-based...applications. For example, specific tissues such as blood cells or neurons can be fluorescently labeled, allowing...divisions, and movements of each of the thousands of cells in the developing zebrafish embryo have been tracked...developing zebrafish embryo has been mapped using single cell RNA sequencing approaches (Harland, 2018). Several...sequenced zebrafish genome. One study profiled 38,731 cells from 694 embryos over multiple stages and reconstructed...reconstructed the patterns of gene activity and cell divisions that coordinate development of the body plan...

...patterning of cells and tissues is fundamental to development. I'm interested in how cells find their way...Position" eBook Here 1. Have a competitive CV You don't necessarily need a Nature/Cell/Science paper. You... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly... What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your aims...human disease" is vague. Instead, consider "A mechanistic understanding of this system will provide the...

...single-cell level. CRISPR libraries CRISPR screening has become a popular tool for probing cellular pathways...barcodes allow researchers to identify specific cells — as well as their descendants. This makes them ... for lineage tracing, among other applications. Cells can be identified using fluorescence, like in the...Figure 1: Schematic of barcoding with a pooled library. Created with BioRender.com. The CellTag libraries...barcoding approach to observe clonal dynamics and cell lineage. Variations of this library allow tracking...mutations that prevent their expression. The knockout cells can be used to identify genes that underlie a particular...genome-wide knockout screens in mouse and human cells, respectively. Last month, the David Root and John...

...Xenopus frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then ...is the smallest photosynthetic cell in existence, but most abundant cell on earth (twice the human biomass...Hawaiian bobtail squid has an extreme number of nerve cells and an unusually high level of RNA recoding in the...using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all (maybe because...total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little bits of hydra can...know that most cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult...organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...

...overexpress YFG in a Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective....person. Animals are diploid except for their germ cells (sets of genomic material are contributed by mother... of their abdomen will be a dark spot. This is meconium (in other words, newborn poop) visible through...

...genes should be ‘read’ by the cellular machinery. This is how most cells in the body contain the same ...visualise changes in histone methylation in living cells. Essentially this reporter construct consists of...recognise lysine 27 methylation. When tested in living cells where methylation of these sites was controlled,...fluorescent reporters of histone methylation in living cells." Journal of the American Chemical Society 126.19...treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon... within an organism. Mechanisms of epigenetic modifications The main mechanism of epigenetic modification...complex of DNA and histone proteins that compacts cellular DNA into chromosomes and reduces the risk of dangerous...

... specific cell-type promoter. Once injected into the brain, the viruses infect all the cells with an inactive...proteins control the flow of electric charges across cell membranes and maintain membrane potential in response...once a certain number of positive ions crosses the cell membrane, a threshold is reached and the neuron ...selective expression of opsin genes in targeted cells makes it possible to control neuronal activity with... This could be overcome by fusing opsin genes to cell-type specific promoters; however, those usually ...drive weak expression of downstream genes. Strong cell-specific promoters, on the other side, are too long...achieve strong opsin expression in specific neuronal cell types then? This is where FLEx enters the scene....

... a common thread at the meeting. The new Somatic Cell Genome Editing program, run through the NIH Common...Improve HDR Efficiency for CRISPR Editing in Human Cells Hands-on tips for repair template design from guest...B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in..., with protocols using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template...to learn much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage..., which alter genomic DNA without the need for cellular HDR.   Having had time to digest my notes, since...a brand new CRISPOR manual to kick off 2018! "Mechanisms of precise genome editing using oligonucleotide...

...mammalian cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this...sequence. That means there are 61 possible tRNAs. Cells however, may not express all 61 of these tRNAs and... are also optimized for expression in mammalian cells. For example, Henry Lester’s lab optimized the C...Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal... cerevisiae (Tuller et. al., 2010). Figure 1: An overview of peptide synthesis. A ribosome interacts...McClain WH (2000) The G·U wobble base pair. EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001... W, Mauro VP (2005) An mRNA-rRNA base-pairing mechanism for translation initiation in eukaryotes. Nat ...

...quality work in cell culture. The problems with cell lines Simply put, the majority of cell line problems...HeLa cells, the first immortalized human cell line. He proved this by examining whether the cell lines...Cancer Cells, Marker-independent identification of glioma-initiating cells. The International Cell Line ...Animal Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen Cell Culture...working cell bank has been made by a research institution (an excellent practice), or maybe cells have been... Outreach Scientist Nick Amiss. Cell culture in the present day Cell culture is a widespread tool used... ability to propagate viruses in cell culture; if there were no cells to infect, there would be no viable...

...the sample (by less than 1%) and recovered after the magnet was removed (Figure 1). At first, Andrew and...visible whether expressed in E. coli, in mammalian cells, or in purified form, and didn’t need any added ...since the initial report in 2023.)     Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli...that could be expressed and purified. Although a ~1% change in signal might not make a very powerful tool...Hayward et al. 2024 under CC-BY license.   Video 1: Fluorescence movie of E. coli expressing MagLOV while...admit they don’t fully understand the physical mechanism, though they have collected a variety of data ...biomedical engineers, join efforts to explore the mechanisms and develop potential applications.  Andrew is...

...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 ...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%...modifications. CRISPR base editing persists through multiple cell divisions, indicating that this method produces ...