We narrowed to 813 results for: cell

...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... 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...optimal sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview...

...knock out, etc. – CRISPR can be used to generate cell line or animal models which have the precise disease...Since CRISPR has been adapted for use in primary cells, many tissue types, and across species, it enables... phenotype. CRISPR can also be used to generate cell line knock-ins of patient mutations which are variants...CRISPR has revolutionized the generation of knockout cell lines which has made this control step extremely...in a cancerous background and treat the parental cell line and the X knockout with your drug(s). If a ...the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the...

...mammalian cells. dDetermined as effective NIR fluorescence in transiently transfected live HeLa cells with ...Near-Infrared Fluorescent Proteins in Mammalian Cells. Cell Chem Biol 24, 758-766 e753 (2017). Baloban, M...developers and users, BV is present in eukaryotic cells as an intermediate of heme metabolism (Shemetov ...to GFP-like probes by supplying a single gene to cells. Still in bacteria, which do not produce BV, an ... to consider effective brightness of NIR FPs in cells, in addition to their molecular brightness (a product...the concentration of BV-containing molecules in a cell and depends on the efficiency of BV incorporation...Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells vs. iRFP713, % d Ref BphP-based FPs miRFP670...

...used to deliver Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid of 2017... coat the virus and help determine the types of cells that it can infect. The VSV-G envelope is widely... lentiviruses to infect many different types of cells. This means researchers who use VSV-G can deliver...deliver their genes of interest to a variety of cells and study their effects. To date, nearly 900 articles...in studies of the secretome, ubiquitin ligase, cellular senescence, autophagy, CRISPR, and so much more...

... the single-cell level can aid in drawing connections between signaling activity and cell phenotype. One...Covert, M.W. (2018). Live-cell measurements of kinase activity in single cells using translocation reporters... One method for monitoring live single-cell kinase activity is FRET, but FRET reporters are challenging...their Kinase Translocation Reporters (KTRs) whose cellular localization serves as a proxy measurement of ... used to measure kinase activity in specific subcellular compartments. Second, because KTRs shuttle between...

...type of cells the viral particles infect. This can direct your biosensor to specific tissues or cell (sub...the caffeine of the cell!)—ATP. There are biosensors for intracellular, extracellular, and even the ADP:...blog we will focus on those which can be applied in cell culture and animal models primarily. Fun fact: Almost...vivo, in tissue preps, in multiple species, and in cell culture models. Some of them have even been expanded... as actinin. When tension is high vs. low in the cell the donor and acceptor of a FRET pair on actinin...many biosensors need to be in specific tissues or cells to monitor their target analyte. This can be difficult... as proxies for a cellular process or directly measure a cellular process. Cellular phenomena such as ...

...proteins in cells. Homeostatic regulation of protein abundance is essential for normal cell function, and...one particular protein in neurons or in any other cell type. However, we cannot distinguish new from old...facilitate the visualization of specific proteins in cells. Tagged protein can easily be overexpressed from...techniques to follow translation in living single cells. However, before CRISPR-Cas9 these techniques were... turnover of specific endogenous proteins in any cell type. Exciting! Many thanks to our guest blogger... how small molecule fluorophores can be used for cell labeling Check out other protein tagging strategies...interests, suspected to interact, are added to a cellular sample. Both antibodies have a short antibody ...

...Translocations in Human Cells Are Generated by Canonical Nonhomologous End-Joining. Molecular Cell 55:829–842 . ...Genome Editing Specificity. Cell 154:1380–1389 . https://doi.org/10.1016/j.cell.2013.08.021 Waters CA, Strande... chromosomal DSBs. It is important to know that cells possess two major repair pathways – Non-Homologous... steps Unlike HDR, NHEJ is active throughout the cell cycle and has a higher capacity for repair, as there...should take less than an hour, thus a population of cells constitutively expressing a targeted Cas9 should...CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video] Pooled CRISPR Libraries Offer Genome-Wide...

...Bewersdorf, J. (2016). Two-colour live-cell nanoscale imaging of intracellular targets. Nature Communications,...opening up new applications.   Figure 1: Live-cell labeling strategies. The green structure represents...organelle interactions. a) Labeling scheme. b) COS-7 cell ER labeled with Halo-Sec61β with SiR-CA (magenta...viral particles with BG-linked antibodies to drive cell type-specific delivery of a viral vector (Strebinger...specialized ligands is more expensive than letting the cells make a fluorescent protein for you. And some ligands...certain probes are not very good at crossing the cell membrane. So, you should investigate your options...Microscopy: A Comparison between HaloTags and SNAP-tags. Cell Chemical Biology, 26(4), 584-592.e6. https://doi.org...

...infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface receptor...antigenic drift.    Starr et al Cell. 2020. https://doi.org/10.1016/j.cell.2020.08.012  Starr et al Science...Library generation and sequencing. From Starr et al Cell., 2020. (B) Mapping antibody-escape mutations with... SARS-CoV-2 spike RBD is displayed on the yeast cell’s surface. The Bloom lab used the library to identify... candidates were validated by IP-MS using HEK293 cell lysate where the antigens would be in their native...immunoprecipitate the entire complex in three different cell lines. The Gräslund lab plans to continue their ...protect proteins from unfavorable interactions in the cell, improving biosynthetic pathway yields by stabilizing...

... are filled with early mitotic cells as well as necrotic or dead cells (Jiang and White-Cooper, 2003)....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...are part of a genetic cascade that determines the cell fate of the dorsal mesoderm in developing Drosophila... and bagpipe: two homeo box genes that determine cell fates in the dorsal mesoderm of Drosophila." Genes...required for oogenesis in Drosophila." The Journal of cell biology 139.7 (1997): 1805-1819. PubMed PMID: 9412474...

...expanded since its adaptation for use in mammalian cells in 2012-2013. Researchers are now using this system...plasmids containing sgRNA sequences into HEK293T cells. The fluorescence intensity was classified into ...is then a simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with...incubation, you assess the level of fluorescence in your cells under a fluorescence microscope. The higher the ...you’ve validated your gRNAs for use in mammalian cells, it’s time to use them in mouse embryos. When researchers...developed as the use of CRISPR/Cas9 in mammalian cells was being explored and it allowed control over each... the mutation will be homozygous for it in every cell; however, if the microinjection was done as the ...

...coordinated by the eukaryotic cell for the execution of many complex cellular behaviors, such as differentiation...toxic intermediate buildup as well as controlling cell fate decisions. By using an orthogonal dCas9 protein...for sequence-specific control of gene expression. Cell, 152(5), 1173-1183. (2013). PubMed. Zalatan, J...Transcriptional Programs with CRISPR RNA Scaffolds. Cell, 160(1), 339-350 (2015). PubMed. Find the Plasmids...crispr/qi/ Plasmids described in  Zalatan et al. Cell  (2015) can be found here: https://www.addgene.org.../article/9593/ Plasmids described in Qi et al. Cell  (2013) can be found here: https://www.addgene.org...or metabolism. Our attempts to manipulate these cellular behaviors often fall short with the generation...

... there is one problem: how to get that into the cells of interest,” Griesbeck said. Because they are chemical...advantage for observing events taking place inside cells that are on the move, such as T lymphocytes. Griesbeck...-binding protein, troponin C (TnC), from muscle cells. They identified a toadfish TnC variant which had...cAMP-responsive element-binding protein (CREB) in living cells. “Now we are making this really good,” Griesbeck...also present challenges in that they are found in cells at much lower concentrations than calcium, requiring...Friedrich et al. Imaging CREB activation in living cells. J Biol Chem. 2010 Jul 23;285(30):23285-95. doi:...understand the connection between molecular- and cellular-level events and behavior. It’s a problem that...

...are commonly found in cells, and without something to block RNase activity upon cell lysis, RNA is degraded...RNases such as RNaseZAP. When harvesting tissues, cells, plants, fungi, or bacteria, keep samples cold and...before RNases had a chance to break down RNA upon cell lysis.  In the late 1970’s, Chirgwin and colleagues...keeping your samples cold until homogenization and cell lysis. This can be challenging depending on your...developed ways to “stabilize” RNA within tissues before cell lysis. These products, namely RNAlater®  from Thermo...solutions that work by inhibiting RNase activity within cells or tissues- they don’t actually chemically stabilize...

...expression and needing to work with a transfectable cell line. 2) They require larger expression vectors... of 4 crRNAs in vitro and when expressed in 293 cells. This allows for a single promoter to drive expression... were used.  With transfection, 6.4% of HEK293T cells had edits at 4 of 4 targets when an array of 4 crRNAs...transfected on a single plasmid. This is versus 2.4% of cells transfected with a pool of plasmids that contained...processing of the crRNA array when expressed in cells, but notice how Zetsche et al reverses the orientation...mouse neurons and in vivo. For these experiments, cells were infected with a 1:1 ratio of two AAVs,  one...RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3), 759-771. PubMed PMID: 26422227. PubMed Central...

...used. The Fujii lab demonstrated this in mammalian cells by expressing dCas9 with an N-terminal 3xFLAG tag...studies (Fujita et al., 2018). In addition to the "in cell" form of this method, in which an engineered DNA-binding...molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii lab also developed another...recombinant CRISPR ribonucleoproteins." Genes to Cells 21.4 (2016): 370-377. PubMed PMID: 26848818. Fujita...‐dC as9 protein for enCh IP analysis." Genes to cells 23.4 (2018): 318-325. PubMed PMID: 29480524. Gilbert...RNA-guided regulation of transcription in eukaryotes." Cell 154.2 (2013): 442-451. PubMed PMID: 23849981. PubMed...of chromatin interactions by biotinylated dCas9." Cell 170.5 (2017): 1028-1043. PubMed PMID: 28841410. ...

...if the cell dies from injury, this protein leaks out into the cytoplasm, migrates out of the cell, and ...Keytruda) causes T-cells to more easily recognize and cause the destruction of cancer cells. Pembrolizumab...cause inflammation, which can cause mesothelial cells to mutate and ultimately form cancerous tumors.  ...resection), systemic chemotherapy to kill cancerous cells that may have spread beyond the point of origin,...successful in killing many (if not all) of the cancer cells that may be left behind after surgery, while being...and binds cells that can begin an immune response. This important biomarker can be used to distinguish between...

...entire proteome of a particular cell type, or a subset based on a cellular compartment or growth stage)....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... DNA and activate a reporter gene in eukaryotic cells, fusion proteins must gain access to the nucleus... the DNA specificity of a prokaryotic repressor. Cell. 1985; 43:729-736. PubMed PMID: 3907859. 4. Fields...

...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...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 ... Recombinant antibodies for marking cell types in the nervous system By Ashley Waldron The mammalian nervous... nervous system is composed of myriad cell types and subtypes, and a familiar challenge for anyone who...

... by ~3,300-fold in Drosophila cells and ~24,000-fold in human HeLa cells as compared to basal expression...origins, Tet systems function well in mammalian cells, and TRE-containing promoters can be used in the...cumate switch system functions well in mammalian cells, and can be used in the repressible manner described...While originally developed for use in mammalian cells, this system has also been implemented in a variety...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system revolutionized...reversible by the addition of quinic acid to Drosophila cells. Work by Wei et al. from Kang Shen’s lab has also.... Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl ...

...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...protein to the existing pool of protein within a cell and may lead to over-expression artifacts. This ...heterotrimeric G protein Gq activation in living cells." BMC biology 9.1 (2011): 32. PubMed PMID: 21619590...activation of heterotrimeric G-proteins in living cells." Science 291.5512 (2001): 2408-2411. PubMed PMID...

...its oxidized derivatives. Cell 152, 5 P1146-1159. https://doi.org/10.1016/j.cell.2013.02.004.  Xu X, Tao...targeted DNA demethylation. Cell Discov 2, 16009. https://doi.org/10.1038/celldisc.2016.9.   ...enzymatic activity cause demethylation?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy...methylate sites blocked by dCas9. Within a few rounds of cell division, the original methylated strand is diluted...dCas9 expression and pLM-CMV-R-Cre. Selection for cells stably and highly expressing these constructs can...can be achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved using...TETs compete with DNMT3 activity in pluripotent cells at thousands of methylated somatic enhancers. Nat...

... used HEK293T cells in our first study, but we're planning to soon expand to other cell types, including...26 of the 29 SARS-CoV-2 viral proteins in human cells and used these viral “bait” proteins to fish for...human proteins that the virus interacts with in cells. Because viruses rely on the help of human proteins...monkeywrench into the virus’ hijacking of human cells. Even better, more than half the drugs hitting these...they have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a fairly...

...different types of human cells, including T cells and hematopoietic stem and progenitor cells. Now with the DAP...to enable multiplex gene editing in a variety of cells and organisms. However, lengthy genetic payloads... over 50% averaging editing efficiency in human cells. MBE with DAP array also exhibited reduced Cas9-... editing, and nuclease editing, in virtually any cell type or organisms of interest with only minimal ...

...expression targeted to specific tissues, cell types or even subcellular regions of a neuron. Optogenetic approaches...approaches excel at discrete cell or tissue-subregion targeting down to the limits of light delivery. ...-wide targeting, as the ligand can reach target cells throughout the body. Controlling Stimulation Some... and chemogenetic tools need to be delivered to cells, commonly through viral delivery by AAV. You can...Further restricted to illuminated region Impacts all cells expressing receptors Invasiveness Mice require...

...potential to form aggregates and may be toxic to the cell. Here comes the SunTag How does SunTag fix these...enables single molecule tracking  within living cells without affecting protein function, thereby creating...attempted single-molecule imaging deep inside the cell, in the nucleus and cytoplasm. Again, they found...SunTag constructs would avoid negative effects on cell physiology. Having seen that mitochondrial tracker...amplification in gene expression and fluorescence imaging.” Cell 159(3) (2014):635-46. PubMed PMID: 25307933. PubMed...time to get enough signal. This workaround risks cellular phototoxicity and eliminates the possibility of...creating a single-molecule reporter of intracellular processes. Initially, Tanenbaum et al. observed some...

...shown to work in plant, Drosophila and mammalian cells, and it may find wide use in biological research...turnover rate. In many cases, however, a GFP-tagged cell line or organism has already been made, and it would...made by accident. After exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. ...spontaneous conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White circles...photoconverted in plant, Drosophila, and mammalian cells. Photoconverted GFP is compatible with other FPs... are you’ve used a GFP derivative to conduct subcellular localization studies or make a reporter construct...this technique may not be appropriate for high-pH cellular compartments. Sattarzadeh et al. have shown that...

...already available today to cure sickle cell anemia in stem cells but this can’t be used in people until... audience discussed how J&J is already modifying cell and animal genomes to test hypotheses of disease...and more about the how to use genome edited stem cells to treat disease. As for me, I continue to be amazed... much we have yet to learn. As Dr. Mello said, “Cells have been in the information age for billions of...

...of fluorescent proteins and their use for imaging cell biology, Michael Davidson’s lab at Florida State...blue, and orange spectra. A comprehensive list of cell markers all tagged with mEmerald. Plasmids to monitor...From F-Actin to mitochondria, from the nucleus to cell membrane, this collection has fluorescently tagged... where your protein of interest localizes in the cell, use these markers as your guide. As Michael Davidson...Fluorescent proteins are] a whole new way of seeing cell biology, seeing what interacts with what, what affects...of many proteins known to localize to specific cellular structures.The Davidson lab even included some...everywhere to get a clearer picture of that intracellular world, even as the Davidson lab prepares to ...