We narrowed to 818 results for: cell
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TypeCollection... optical switch tools to measure and manipulate cells and their biomolecular processes....to blue light and allows cations to flow into the cell. Variants of channelrhodopsin have been engineered...improve the expression and membrane targeting in host cells. Other microbial opsins, such as the Halorhodopsins...actively move cations or anions into or out of the cell. When using these opsins as a tool to control neuronal...plasmids that use light to control a protein or cellular activity. For example, search "Cdc42", "transcription...
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Tetracycline Inducible Expression
TypeCollection...Plasmids Collection Cre-lox Plasmids Collection Stem Cell Research Plasmids Collection Tetracycline Off Tetracycline...expression of the transactivator proteins in mammalian cells or other species by codon optimization of their ...gene of interest from two separate vectors. Stable cell lines or transgenic animals can be made to achieve... and ensures that all transfected or transduced cells express both components in the same relative ratio...derivatives can be found in Krueger et al., 2004. Some cell culture sera may contain tetracycline or its derivatives...expression of mouse Oct4, Sox2, Klf4, and Myc for iPS cell generation Rudolf Jaenisch 51543 FUW-tetO-hOKMS ...expression of human Oct4, Sox2, Klf4, and Myc for iPS cell generation Tarjei Mikkelsen 172115 PB-TO-hNGN2 Piggybac... -
Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]
TypeBlog Post...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... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog Post... 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... -
RNA Extraction Without A Kit
TypeBlog Post...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... -
Multiplex Genome Editing with CRISPR-Cpf1
TypeBlog Post...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... -
Isolating Genomic Regions of Interest with the CRISPR System
TypeBlog Post...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. ... -
Mesothelioma - Causes, Symptoms, and Treatment
TypeBlog Post...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... -
Tips for Screening with Yeast Two Hybrid Systems
TypeBlog Post...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... -
AAV Purification by Iodixanol Gradient Ultracentrifugation
TypeProtocol...suitable for virus purification and isolation of cells, organelles, lipoproteins, and macromolecules. Importantly..., iodixanol is inert and non-toxic to mammalian cells. Therefore, and unlike other density gradient media...