...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...purpose and mechanism of polyadenylation vary across cell types, but polyadenylation generally serves to promote...additionally act as a quality control mechanism to rid the cell of mis-folded RNAs. Eukaryotic termination and polyadenylation...efficient rabbit beta-globin mRNA 3' end formation. Cell. 1987. PMID: 3568131.  Hager S, Frame FM, Collins... thought that poly(A)s are used to control the cellular concentration of regulatory RNAs and may additionally...signal by using in vitro analyses. Molecular and Cellular Biology. 1992. PMID: 1333042. PubMed Central PMCID...

...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...complex of DNA and histone proteins that compacts cellular DNA into chromosomes and reduces the risk of dangerous...the more compacted it becomes. This prevents the cellular machinery from physically interacting with the...

...school at MIT. “I read some of the seminal papers on cell cycle discovery from Lee Hartwell and Paul Nurse...genetic screen to identify the factors that regulate cell division. And that really got me hooked on genetics...high-resolution tracking of more than 1 million cancer cells under drug treatment.” Stegmeier especially finds...

...graduate student studying plant development and cell-cell communication in the Nimchuk Lab in the Biology...untransformed seedlings or callus (undifferentiated plant cell culture), which is great for a wide variety of applications...

...Fluorescence Activated Cell Sorting (FACS), allowing for enrichment of successfully targeted cells that can be used...investigating genetic mechanisms related to morphogenesis, cell fate specification, and human disease.     Sun,...

...fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys at a specific developmental...genes of interest may not be expressed in the tissue/cell line you are working with. Development-specific...10.7554/eLife.67667 Springer, S. P. Gene Traps. Plant Cell. (2000) 12(7): 1007-1020. DOI: 10.1105/tpc.12.7.1007...

...two-photon microscopy. Cell, 18, 3408 - 3425.e29. (2022). https://doi.org/10.1016/j.cell.2022.07.013.   ...voltage fluctuations. Image credit: Liu Z, et al. Cell. 2022.  Find JEDI-2p viral preps here (and here...specific for human c-myc proto-oncogene product. Mol Cell Biol. Dec;5(12):3610-6.(1985). doi: 10.1128/mcb.5.12.3610...

...Mammalian cells provide scientists with the means to study biological processes on the cellular level instead...with tissue culture The cultivation of mammalian cells in the lab, or tissue culture as it is commonly ...

...to detect dead or dying cells so that you can base your analysis on living cells only (for example, propidium...This tends to be most problematic when staining cells or tissues that are the same species as your primary...propidium iodide only stains dead cells). Compensation controls address the spectral overlap that occurs...Quantitative Fluorescence Microscopy.” Journal of Cell Biology, vol. 185, no. 7, Rockefeller University...

... edit A → Y (Y = T or C) in 67.17% of screening cells (Tong, Wang, et al., 2023). Based on this promising...Cas9 nicks the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template... efficient A•T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4), 638–650...programmable T-to-G and T-to-C base editing. Molecular Cell, 84(7), 1257-1270.e6. https://doi.org/10.1016/j....efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547. https:...the base editor. Black arrows indicate natural cellular processes. Created with BioRender.com. The...

...two-photon microscopy. Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi: 10.1016/j.cell.2022.07.013 pYtags...Calcium Indicator. Front Cell Dev Biol. 2022 Jun 29;10:880107. doi: 10.3389/fcell.2022.880107. KRaION potassium...sensor relies on a change in localization to the cell membrane and not a direct change in the fluorescent...of receptor tyrosine kinase signaling in living cells. bioRxiv 2022.08.13.503850; doi: 10.1101/2022.08.13.503850...better-matched to the potassium concentrations in cells than existing sensors. Using genome mining and structure-guided...fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent...inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled receptors....

...use electro-competent cells instead of the more common chemically-competent cells. You should perform, ... manufacturer’s instructions for your competent cells. For most standard cloning, you can transform 1-...1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using much less total...colonies, you might want to use higher competency cells. Additionally, if your final product is going to...

...high light doses to the cells. Thin static samples Ex: Fixed monolayers of cells or thin (<15 μm) tissue...1). Thin dynamic samples Ex: monolayers of live cells with fluorescently labelled, motile structures These... 10s of nanometers. Thick dynamic samples Ex: 3D cell cultures, small model embryos >15 μm Thsee are some...Total internal reflection fluorescence microscopy in cell biology. Methods Enzymol 361, 1-33 (2003). PubMed...Richardson DS, Lichtman JW. Clarifying Tissue Clearing. Cell 162, 246-257 (2015). PubMed PMID: 26186186.  Additional...Biological Imaging and a Lecturer on Molecular and Cellular Biology at Harvard University. No matter whether...Biological Imaging and a Lecturer on Molecular and Cellular Biology at Harvard University.     Additional ...

... acid (DAP), a component of the cell wall. Once in the recipient cell, the CRISPRi DNA (purple) flanked... is propagated during normal DNA replication and cell division. Mobile-CRISPRi does not disrupt gene function...mobilize these plasmids for transfer to recipient cells during a tri-parental mating scheme. Mobile-CRISPRi...

... in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct NGG PAM NGA PAM NGC PAM NGT... percent editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages...

... make it difficult to properly gate your desired cell population. By compensating between fluorescent ... your fluorescent markers are staining for. If a cell is positive for both markers, it will show up as...Figure 3: Flow plot for a double positive sample. The cell population in the top right corner of the plot is...learning! Ashlyn has a research background in CD8+ T cells and cancer immunotherapies. She is currently a research...

... Typically, Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation...DNA. This bias can be counteracted by growing the cells on solid plates, where each transformant forms its...own colony, uninfluenced by the growth of other cells. For libraries where the plasmids are all very similar...

...reliably detect HMOs. Fatima’s first design is a whole cell biosensor with two components - an enzyme that breaks...binding results in an increase in fluorescence of cells that actively produce the HMOs. Fatima has demonstrated...out Fatima's recent publication on her HMO work in Cell Chemical Biology! Additional Resources on the Addgene...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their...proposes using this sensor to detect HMOs found on cellular surfaces. After incubating the sensor with bacteria...

...to supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved...Imaging Center at UCSF. A common requirement for live cell imaging experiments is the ability to follow multiple...

...would continue on with my academic dream of studying cell division. However, after 6 years in the business...Cambridge University where he studied mechanisms of cell division. Recently Seán got the entrepreneurial ...information on ELISA troubleshooting, ELISA kits and cell death assays. You can find Seán on twitter @Macfhearraigh...