...recombination varies for different constructs and cell types. Second, Cre may induce recombination at pseudo...standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding domain-fusion...recombination efficiency. When expressed in neuronal cells, each line showed broad recombinase-dependent RFP...these recombinases were also active in mammalian cells. In experiments, B3 and KD were shown to be functional...functional in cultured Chinese hamster ovary (CHO) cells, as well as in cortical neurons in layer 2/3 of ... damage induced by Cre recombinase in mammalian cells. Loonstra A, Voojis M, Beverloo HB, Al Allak B, ...recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity regulation. Heidmann...

...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...of many proteins known to localize to specific cellular structures.The Davidson lab even included some... 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...everywhere to get a clearer picture of that intracellular world, even as the Davidson lab prepares to ...

...eight millimeters – not single cells (yet) but small clusters of cells. Looking ahead What’s next, you...fluorescent proteins to observe what’s happening inside cells for at least a couple of decades. Green is the classic...and tracking of proteins, organelles, and living cells in a spatiotemporal manner, which is not possible... vivo. “One of the reasons we can’t localize one cell at a depth of one centimeter with fluorescence is...on the other hand, are larger than the size of a cell, so they scatter much less. As a result, ultrasound...brighter, more photostable, smaller, and less toxic to cells. They’d like to make near-infrared fluorescent proteins...

...identification of potential targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens...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...

...time to get enough signal. This workaround risks cellular phototoxicity and eliminates the possibility of...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...creating a single-molecule reporter of intracellular processes. Initially, Tanenbaum et al. observed some...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...

... disease, cell type or drug treatment can browse through screens performed in relevant cell lines or conditions... currently houses over 890 CRISPR screens in 629 cell lines, each manually curated and annotated from ...feasibility and power of CRISPR-based library screens in cell lines from humans and other species. By accurately... screens performed in 26 different breast cancer cell lines. Screenshot updated 2024. Each screen ...screen is captured by BioGRID curators, including the cell line/tissue type, enzyme system, sgRNA library and...example phenotype screens with read-outs other than cell viability. The following video provides an in-depth...

...to reproduce itself as it must to survive within cells. The replicons of plasmids are generally different...’s sake, we've ignored those used in eukaryotic cells and viruses and focused only on those found in bacteria...example, the pMB1 ori maintains about 20 copies per cell, while pUC – which differs by only two mutations...mutations – will produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard ...positives? Will you have only one plasmid type in your cells at a time? Do you want to make a lot of your plasmid... stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies streaked...bacteria have just reached stationary phase, but the cells have not started to die off. Have any questions...

...mCherry for FACS sorting of the weaponized T-cells (CAR-T cells) - and then delivering them to a tumor - ...effectively as standard CAR-T cells, but it reduces the off-target effects caused by T cell migration.  CAR expression...reducing the on-target off-tumor activity of CAR–T cells was recently devised by the Yingxiao Wang lab.  ...above can be made for expressing your favorite T-cell receptor, or perhaps any other tool that can benefit...  Wu Y, et. al. Control of the activity of CAR-T cells within tumours via focused ultrasound. Nat Biomed...

...plant protein TIR1 in non-plant cells, which interacts with the cells’ endogenous ubiquitin ligase, the...translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al., 2016...based method to tag endogenous proteins in HCT116 cells with mAID. Using this method, the lab was able to... protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that...the target sites adjacent to an NG PAM in human cells. The increased versatility of the engineered SpCas9... just a 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised of thousands...

...non-growing cells are orange, slow growing cells are orange-green, and fast growing cells are green. A...fluorescence. Cells expressing the library are then sorted by fluorescence-activated cell sorting into ...population. Last, Bernach will simultaneously monitor cell cycle and growth rate using TIMER and an unstable...reporter under the control of promoters that control cell cycle gene expression.  Additional Resources on...

...’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new...spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find an app that could...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...tools are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution...educational section. This section holds the Gibco Cell culture manual and basic instructional videos. The... Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England BioLabs...alternative, similar apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme...

...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...

... Stem Cells Using Type I CRISPR-Cas. Molecular Cell, 74(5): 936-950.e5 (2019). 10.1016/j.molcel.2019.03.014...a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016/j.cell.2015.09.038 Harrington, B., ... Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral RNA...cleavage and is significantly less toxic to mammalian cells than Cas13 (likely due to the lack of collateral...

...level, in the case of some public institutions) to cancel plans to raise salaries. What are institutions ...expecting a salary increase have had that increase cancelled. Rounding out the remainder, 2% of postdocs who... are continuing with their plans, and 17 have cancelled their plans. 2 of those raising - the University...Illinois and the University of Michigan - initially cancelled their plans, but have since reversed their position...announced that they were reversing their plans to cancel raises). A further 9 institutions, whose plans ... In addition, PIs at some of the institutions cancelling plans are going ahead with raises anyway, including...injunction, as we started to hear of institutions cancelling plans to raise salaries, the only postdocs we...

...indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured. Archon1 ... it. Using a robotic picking approach to select cells with variant characteristics they wanted, the team...offers a way to spatially and temporally control cellular processes in vivo. Many photoactive proteins exist...approach to control protein-protein interactions in cells because the protein binders can be customized for...photo-reversible protein-protein interactions can control subcellular localization processes in vivo. They deposited...TOM20 (pTriEX Tom20 mVenus BoPD). In dark adapted cells expressing these plasmids, tgRFP fluorescence localizes...

...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...

...in transfected eukaryotic cells, allowing for them to be used as intracellular inhibitors.  Aptamers are...the kids say) affinity reagents deserved to be celebrated, there are many other amazing affinity reagents...small enough to be expressed in bacterial and other cell systems. Since the majority of these alternative...the constant region. Nonspecific binding, caused cell receptors for the constant region, is also reduced..., peptides, amino acids, drugs, metal ions, and cells. They can be used either with or as fluorophores... may sound. Validated antibodies are still an excellent option for many applications, and frankly, if ...

...producer cell line, such as HEK-293T, production of viral particles, and transduction of a target cell line...re derived from viruses that can infect people, cells, and animals. Some of these viruses are completely...While the virus can still enter (and thus infect) cells, this safety feature prevents the virus from replicating...replicating and spreading in non-target cells, including yourself as the researcher.   Figure 1:...vector carries an shRNA to a gene essential for cellular function, this would also be a higher risk vector... the procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-...

...year and how these plasmids have been used to accelerate COVID-19 research in other labs. Spike protein...depends on its spike protein to bind ACE2, the host cellular receptor. To more easily study the spike variants... investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids express these...for SARS-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their search, they found...packaging) and contain an IRES-Puro marker for stable cell clone selection. For more information on these plasmids... during the COVID-19 pandemic. Our mission to accelerate research through sharing reagents has never been...Bajic G, Harrison SC, Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-...

... be introduced into host cells, such as bacterial, yeast, or mammalian cells, for antibody production ...Nanobodies and recombinant binders in cell biology. Journal of Cell Biology 209:633–644 . https://doi.org...animals and bleed dates and even clonal hybridoma cell lines can produce more than one mAb (Bradbury et...plasmid or from an integrated sequence in a stable cell line. The genes encode the heavy chain and light...antibody protein binds an antigen. Image from Fvasconcellos.     How are Recombinant Antibodies Made...