We narrowed to 1,027 results for: ell

...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 ...Viruses are simple: a genome packaged in a protein shell (Taylor, 2014). They’re so simple that we can’t ...While all viruses have a genome inside a protein shell known as a capsid, some also have a lipid bilayer...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...

...guides, short pieces of RNA that bind to a cell’s DNA, telling the Cas machinery where to cut.      ...Biomedical research has a long and well-documented history of racial bias, which still impacts and influences...ancestry and germline sequence variation in cancer cell lines took them down unexpected paths. Though this...researchers do large-scale CRISPR screens in cancer cell lines, looking to uncover genetic dependencies of...to see if they could identify the ancestry of the cell lines (European, East Asian, Indigenous American...or South Asian) used in the screens based on the cells’ germline genetic variations. They then wanted to...with each ancestry group. Since about 90% of the cell lines used in DepMap CRISPR screens are of either...

...entire proteome of a particular cell type, or a subset based on a cellular compartment or growth stage)....strain for screening), or arrayed on a 96 well plate where each well contains a defined prey fusion.  The ...El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi, R., Ledent...extensively to screen for molecular interactions in the cell, including protein-protein, protein-DNA and protein-RNA...field of eukaryotic transcriptional activation and cell biology. During this period, proteins were successfully...The reporter gene then allows detection of those cells where binding between the proteins of interest has...alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the problem is that your proteins are not post-translationally...

...genome-wide interrogation of the genes required for cellular processes. For a successful screen, it is necessary... on the method used to introduce the gRNAs into cells. Most CRISPR pooled libraries have been created ...the chance of transducing a only a single gRNA per cell. The viral DNA containing the gRNA then integrates...next-generation sequencing of the genomic DNA. However, some cell types are poorly transduced by lentivirus, and thus...CRISPR libraries was not possible. Primary murine T cells cannot be transduced by lentivirus and have been...library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express Cas9 and screen...CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction, ...

... are filled with early mitotic cells as well as necrotic or dead cells (Jiang and White-Cooper, 2003)....a key role in proper limb and digit formation as well as neural development. Specifically, SHH is a morphogen...Ivics et al 1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene called "...the fly actomyosin cytoskeleton that regulates cellular contraction. This network is composed of actin...interact with each other to generate forces that cause cells and muscles to contract depending on the context...context. Contraction is critical for processes such as cell movement/motility, development, cytokinesis, and...scientists who discovered comr thought the sperm cells in mutant flies looked like a “whole bunch of cookie...

... preparations are added to cells in culture. Three to five days later, cell cultures are inspected for...a new vial of early passage cells. Approximately two weeks post-thaw, cell culture supernatant is tested...section for images).     Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre... (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445...preparations ("-LV” catalog items) are produced in 293T cells using the 2nd generation psPAX2 and pMD2.G packaging...packaging system developed by the Trono lab. Cell culture medium (OptiPro SFM) containing lentivirus is first...(PEG) followed by centrifugation. Precipitated pellets containing viral particles are then resuspended...

...Addgene to advance their research. Louis Bengyella Dr. Bengyella’s work has focused on elucidating the molecular...his future work. Dr. Bengyella would like to use the expertise he’s developed as well as CRISPR-based gene...chimeric cellulases that enable E. coli and Citrobacter freundii to generate sugars from cellulosic biomass...our first two plasmid grant awardees: Drs Louis Bengyella and Kwabena O. Duedu, both from the University...in Ghana. Read on to learn more about doctors Bengyella and Duedu and how they plan to use plasmids from... microbial pathogens. This work has given Dr. Bengwella expertise in using bioinformatic and proteomic...development of uterine tumors. According to Dr. Bengyella, this class of tumors is “a major public health...

...Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing. Cell Stem Cell 15:27–30 . https://...protein, in order to mitigate the ability of the cell to use an alternative ATG downstream of the annotated...process, and usually involves the need for single cell cloning and subsequent screening for successful ...standard requires not one but two rounds of single cell cloning – as a control, one should revert the edit...passenger variant that came along with the single cell clone (although this is rarely done). When targeting...can affect the accuracy of a predictive score, as well as dynamic aspects of the target (e.g. accessibility...levels of modification. Whole-genome sequencing of cells modified by CRISPR indicates that the consequences...

...editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652.e29. https...efficient prime editors. Cell, 186(18), 3983-4002.e26. https://doi.org/10.1016/j.cell.2023.07.039 Lin, Q.,...correct only a minority of those variants in most cell types. But prime editing, a CRISPR technique developed...unedited DNA. The heteroduplex is resolved by the cell’s mismatch repair system, and more recent prime editing...than prime editing. But for positions that aren’t well positioned within the editing window, prime editing...editing demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations...type of edit, genomic context, delivery strategy, cell type, and goal of the experiment can all impose ...

...analyzed gDNA from a few thousand cells from single wells of a 96-well plate without prior quantification...was contributed by Scott Findlay, a Postdoctoral Fellow at the University of Alberta. If you’re like many...physically isolated partitions (such as droplets or wells on a chip), such that most partitions will contain...are many thousands of partitions, statistical modelling can be used to determine the actual number of ...and reverse primer to amplify the target locus, as well as a reference probe designed to bind at an unedited...to quantify rates of mutation in a bulk or single cell-derived genomic DNA sample as part of a precision... that they are easily able to distinguish single-cell derived samples with mono-allelic mutations versus...

...Deisseroth 83895 AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Gordon Fishell   Optogenetics Optogenetics continues...Cepko 83898 AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 Gordon Fishell   Chemogenetics Chemogenetics offers...pAAV-hDlx-GiDREADD-dTomato-Fishell-5 83897 AAVrg pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Controls Plasmid...pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg pAAV-mDlx-GCaMP6f-Fishell-2 ... results, but also confirm expression in target cells. While the various AAV serotypes can be used to ...many of our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...offers noninvasive methods for manipulating cell activity, particularly in the brain, and holds promise in...

...gets interrupted as shown in cell A, and thus will not complement the cell's β-galactosidase mutation. An...therefore the cell will have a functional β-galactosidase enzyme through α-complementation (cell B). As shown... plasmid-containing cells without using antibiotic resistance. These rely on cell lines susceptible to...without which the cells cannot reproduce to form colonies (used to complement auxotrophic cell lines), or genes...pUC18 and pUC19 Let’s begin at the beginning. The well-characterized bacterial lac operon contains a gene...called the α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a...screening process. It does not select only those cells that have taken up a plasmid and thus should be ...

...building-blocks of living cells, rapidly forming covalent bonds to specific partners under cell-friendly conditions... can achieve, but are simple to pick up and work well the first time. But biology is complicated — so ...using the reactive groups naturally present in a cell? By analogy to Click Chemistry, Click Biology can...the diverse kinds of proteins that one finds in cells. High yield of reaction — generating product without...reduced; in the ER and Golgi apparatus and outside the cell, most cysteines are oxidized in disulfide bonds....temperature and on ice, when we have cracked open the cells. The tools should react in all of these situations... for connecting proteins in the test tube or in cells. There are also SNAP-tag and HaloTag modules, which...

...proteins enable the observation of proteins in living cells and organisms. Both components of the chimera are...However, these may not reflect the situation in cells. Therefore, the localization of critical fusions...currently the standard for assessing oligomerization in cells. In addition to measuring stickiness, several researchers...fluorescent proteins that behave as monomers in mammalian cells include: mTurquoise2, mEGFP, mNeonGreen, mScarlet...figure 1). In fact, for several FRET biosensors (yellow cameleon 3.60, EPAC, and Galphai), the C-terminus...of the fluorescent protein into the POI can work well since the N- and C-terminal residues of the fluorescent...protein to the existing pool of protein within a cell and may lead to over-expression artifacts. This ...

...epigenome editing. Cell, 184(9), 2503–2519.e17. doi: https://doi.org/10.1016/j.cell.2021.03.025.    Recombinant...tissue is identifying the cells you are studying. Antibodies can help reveal cellular identities by targeting...Trimmer’s lab (Figure 5) as well as antibodies for identifying other cell types in the nervous system!..., particle orientation distribution and Fourier-shell correlation curve (D), and the reconstructed 3D ...particles (eVLPs) offers an option to transduce cells directly with ribonucleoprotein cargo. eVLPs contain... will only be transiently present in transduced cells, limiting off-target editing. Cargo size limits ...site-specific integration of large genes in mammalian cells via continuously evolved recombinases and prime ...

...transfected into a packaging cell line, usually a derivative of 293T. Packaging cells are incubated and after...fine-tuning selection and expression in your target cell line, researchers often find themselves faced with...impact the transfection efficiency of the packaging cell line and thereby affect the titer of the virus. ...layer flask will require some initial optimization; cell number, media volume, and plasmid concentrations...larger vessel to the 10 cm2 dish and adjusting the cell number, plasmid concentration, and experimental ... consideration when titering a lentivirus is the cell line that is used for titering. Typically researchers...researchers will choose a relatively permissive cell line for titering in order to obtain the best titer possible...

... stem cells. The lab will use plasmids from Addgene to construct NFIX expression vectors as well as reporters...the physiological and pathological roles of human cell glycans. Accordingly, she joined efforts with the...aspect of immunology, participating in events such as cell development, recognition and migration, the immune... is not surprising. However, the molecular and cellular mechanisms behind the immune defects in CDG patients...plasmids to immortalize CDG patient-derived immune cell lines and use these lines to study how immune processes...and other pathological conditions. References: Monticelli, M., Ferro, T., Jaeken, J., dos Reis Ferreira...McMurray is an Associate Professor in the Department of Cell and Developmental Biology at the University of Colorado...

... expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown... complex. Not performed in a native cell context. For AP-MS, cells are lysed before the bait protein is...and interacting proteins in mammalian cells. The Journal of cell biology. PubMed PMID: 22412018. PubMed... it allows for the validation of a binary PPI as well as the identification of additional interacting ...against PPIs that aren't stable outside the native cell context. The Split-BioID method Enter Split-BioID...protein interactions. However, FRET generally only tells you if two proteins interact and cannot be used ...tagging of proteins in both BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein...

... require minimal cellular perturbation. Also, this method requires live, intact cells; the sequencing ...Synthetic recombinase-based state machines in living cells” (1), into a more straight-forward, concise explanation...mutations, and environmental conditions drive certain cellular behaviors, but little is known about the order... may be used to better understand and engineer cellular processes that are influenced by temporally ordered... one final state, then the state of the RSM will tell you precisely what inputs were added to the system... our paper (1), we allowed individual bacterial cells to form colonies on a plate and then we performed..., but more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The...

...metabolite in a cell. The only way to do that at scale right now is to grow up a bunch of cells, sacrifice ...capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks Foundation...transistor-like digital genetic switches that enable cellular computing. These logic gates built from transcriptors...available at Addgene as a BIOFAB kit comprised of well-characterized bacterial transcription and translation...as an operating system for engineering an entire cell. Typically when people use plasmids, they are trying... you could imagine engineering the entirety of a cell. That's a big long-term technical challenge. We ...That's central to everything that goes on inside a cell. Until we get a handle on precise, forward engineering...