We narrowed to 595 results for: cel.2

...PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's user guide." Trends in cell biology...imaging in a variety of cellular compartments including the secretory pathway (Figure 2), the inner membrane...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...numerous cell biology assays. However, there has been a pervasive view that most FPs in a cell will fold...fluorescent fusion proteins in cell biology."Current Protocols in Cell Biology (2005): 21-4. Pubmed PMID...Fluorescent proteins in cellular organelles: serious pitfalls and some solutions." DNA and cell biology 32.11 (...-fusions on quantitative imaging experiments and cell physiology in general. Pitfalls of Fluorescent Protein...

...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...untreated cells—but can we be certain? Do we know whether the plasmid delivered into the cells successfully...amount of DNA transfected into the cells and the ability of the cells to express the protein.   In analysis...Gene X in backbone Y) was transfected into human cells. From this result one could simply conclude that...didn’t work, as the expression level of Gene X in cells treated with Plasmid A is similar to the expression...

...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Figure 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...epithelial cells, liver, CNS (limited penetration), retina, hematopoietic cells, and immune cells. B) AAV11...Sanchez, P. S., & Rodriguez-Fraticelli, A. E. (2025). Pre-existing stem cell heterogeneity dictates clonal...the acquisition of leukemic driver mutations. Cell Stem Cell, 32(4), 564–580.e6. https://doi.org/10.1016...therapy and ex vivo cell engineering applications and is especially well-suited for lung cells via airway administration...

...AAV8 for kidney cells — can help improve infection rates in your experiment. Figure 2: Visual representation...handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires a less involved setup when working..., 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández, A., Low, S. H., François, A.,...same: Deliver CRISPR library to cells of interest. Select for cells of interest (positive or negative...infect specific cell or tissue types. Using serotypes that have a known tropism for your cells of interest... a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing cells for long...mice are involved. Ex vivo starts with transducing cells with the CRISPR library in vitro before transplanting...

...and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures GFP is limited in its...proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays a critical...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...ability to interact with protein partners within the cell. However, antibodies can have low affinity for these... generally bright, stable and well tolerated by cells. They can be used for protein localization, isolation...trigger protein aggregation which is toxic for the cells and affects the labeling of small structures like...team, an ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with...

...Quotes taken directly from participants (summarized in 2) touched upon many problems in this area: “ [The] ...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. McDowell, Gary S., et al. "Shaping the Future of ...A fair deal for PhD students and postdocs." Elife 2 (2013): e01139. Pubmed PMID: 24137543. Pubmed Central...following panel members: Jonathan Dinman, Chair of the Cell Biology and Molecular Genetics Department at University...

... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...Biotechnology. https://doi.org/10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, ... systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016/j.cell.2023.03.034 Resources...Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016/J.CELL.2013.07.018 Anzalone, A....epigenetic manipulation of prime editing. Cell. https://doi.org/10.1016/j.cell.2024.03.020 Mathis, N., Allam, A... balance. Molecular Cell, 81(10), 2216-2230.e10. https://doi.org/10.1016/J.MOLCEL.2021.03.032 Yu, G., ...

...channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to easily detect opsin-expressing cells. The resulting.... "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit ...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents... specific cell-type promoter. Once injected into the brain, the viruses infect all the cells with an inactive...energy conversion or the induction of intra- or intercellular signaling pathways (Ernst et al., 2014). Microbial...proteins control the flow of electric charges across cell membranes and maintain membrane potential in response...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET... primary cells. The authors used this platform to assay dozens of GPCRs across many cell types in...biosensor platform. Cell, 187(6), 1527–1546.e25. doi: https://doi.org/10.1016/j.cell.2024.01.028. New...Prime editing (PE), initially developed in human cells, has been uniquely challenging to apply in dicotyledonous...labeling approach using these constructs in HEK 293 cells (Smith et al., 2024). They then used single-molecule...

...portion of our careers. In that time we may have only 2-3 formal supervisors that will provide guidance and...be challenging – is advice and guidance from only 2 people enough?  For many years I’ve been organizing...Apparently, a good Jedi does not necessarily an excellent mentor make. Are your current mentors/advisors...What Kind of Mentor Do I Want To Be?” In this excellent essay Acclimatrix, an assistant professor at a...

...(you generally don’t want more tag than protein), 2-3 tags is increasingly common. Tandem affinity purification...TEV cleavage site (more on that in a moment), and 2 ProtA IgG-binding domains. TAP has since come to encompass...found in this paper (available at Addgene).   Table 2: Protease recognition sites commonly used with tags...not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor... that your reference genes are suitable for your cell lines and/or tissue types.  Please do note my oh-so-subtle...reference genes to save on plate space. For many cell lines and tissue types, appropriate reference genes...

...untagged expression of SARS-CoV-2 proteins in bacteria, yeast, and mammalian cells. We teamed up with Ginkgo...approximately 3,000 constructs to support SARS-CoV-2 research.  Browse the entire Ginkgo Bioworks COVID...organizations to share materials more broadly to accelerate COVID-19 research: Sinai Health System                                     ...

...literature? Very often the data of interest are in 1 or 2 figures from one paper and then another figure in ...ReFigure by creating ReFigures and answering this 2 minute survey! You can also find ReFigure on Twitter... remember what happened when drug X was added to cell line Y, or the phenotype of a mouse with a specific...comparison of the same experiment from different cell lines/model systems ReFigure allows you to make...

...you look at the dot plot in Figure 2, you might assume that all cell populations are positive for both ... the case of Figure 2, one population is B- and another two are B+. Our A-B+ cells are in the lower right...right corner.      Figure 2: This flow plot has two populations that are B+ and one population...-B+ cells, and A+B+ cells, but no cells that are A+B-.      Figure 3: It's often easier to identify...population of B cells and dendritic cells (DCs) in the lower middle right. DCs and B cells are HLA-DR+ and...single cells in a population. It can be used to measure protein expression, identify rare cells, or even...looking for cells in a sample that are negative for marker A and positive for marker B, or A-B+ cells. If you...

...starting day 2 after transfection.     Fig: 2: Expressing mARGAna in MDA-MB-231 cells. (a) Plasmid...expression. When engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...white inclusions in cells with GVs via phase contrast microscopy, GVs visible in cells via TEM, and purified... heterogeneity required single cell screening to isolate a clonal cell line capable of producing ultrasound...expression homogeneity, as the cells could be batch-sorted to isolate a polyclonal cell line with strong gas vesicle...transient transfection in rtTA+ cell lines (such as HEK293-TETOn cells), or they can also be integrated...

...Epub 2016 May 2. PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan... be stable in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast...unable to show TtAgo-mediated cleavage in mammalian cells, likely due to high temperature requirements. To...plasmid and genomic DNA in a variety of mammalian cell lines, with an efficiency similar to that of Cas9...

... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ...incubated at stationary phase for too long, the cells in your culture will no longer be viable. The time...graduate student at Stanford University. She studies cell fate maintenance in the stomatal lineage and has...

...daughter cell, instead of partitioning each type of plasmid into a separate daughter cells (Figure 2) (Ebersbach...to plasmid loss (Diaz et al., 2015).  Figure 2: The partitioning problem. For low copy plasmids...Plasmids. Microbiology and Molecular Biology Reviews 62(2): 434–464. Pubmed PMID: 9618448 Diaz R, Rech J, Bouet...copy number in the cell is high, but allows replication when copy number in the cell falls too low (Novcik...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...eukaryotic cells). The partition NTPase is recruited and “walks” each plasmid to a separate daughter cell. When...from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...

...vector:insert ratio is used in the DpnI digestion and 2 µL of digest product are used in the transformation...Biology 38 (2016): 145-154. PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified...properly. Directly transform chemically competent cells with the digested mixture. There is no need for ...purification before transformation. After transforming cells, you should perform colony PCR with vector specific...interested in protein science.     References: 1. Celie, Patrick HN, Annabel HA Parret, and Anastassis Perrakis...

... than control, 3; same as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice...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 ...

... environmental sustainability in mind (Table 2). Table 2: Sustainability features highlighted in plasmid...plasmid DNA from unwanted cellular “debris”; and then clean-up the plasmid DNA (Figure 2). For most kits, the...together — no mixing and matching!   Figure 2: Plasmid isolation workflows are similar across kits...the steps for lysing cells and removing cellular debris look very similar. However, keep in mind that ...relative term — even armed with the highest-competency cells you will still struggle to get transformants if ...quantity) – Depending on the experiment, mammalian cell transfections can require quite large amounts of...your bacteria to release the plasmid, separate the cell debris from the plasmid, and then clean-up the plasmid...

...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...Xenopus frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then ...is the smallest photosynthetic cell in existence, but most abundant cell on earth (twice the human biomass...only 1,700 genes total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little...Mormyridae (Jason Gallant) Weakly electric fish are excellent systems to study for convergent evolution because...

...into the several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, ...mounting sample on slides, and confocal   Figure 2: High-level overview of an indirect immunofluorescence...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment...interest live within a cell,” “Does this disease change the architecture of my cells,” or “How does this ...IF), is an immunoassay that brings to light the cellular world. The technique allows you to ask questions...this mutation impact the types of cells found in my tissue.” It is based on the same principles as other ... visualize a target of interest within an intact cell, tissue, or, in some cases, a whole organism. But...

...Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Find...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...are capable of cleaving genomic DNA in mammalian cells and zebrafish embryos, and they can be used to modify... adapted for targeted RNA cleavage in mammalian cells. The REPAIR (RNA Editing for Programmable A to I...reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol 34:863–868 . https://doi.org/10.1038...specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol 34:869–874 . https://doi.org/10.1038...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor... of all of the mRNA bound-ribosomes in a cell. When cellular mRNAs are digested with ribonucleases, ribosomes...Regulating translation is key to cellular function, especially during development or stress. With ribosome...explore the complexity of translational control in cellular physiology. Ribosome profiling Ribosome profiling...the effects of 5’ UTR variants in translation of cell cycle protein Emi1, showing that ~80% of transcripts...powerful tools for studying translation in various cell types and physiological states. References 1. Ingolia...

...Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence...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...

...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...useful addition to the toolkit for visualizing cellular processes in C. elegans: mScarlet3 for applications...biologically relevant forces at focal adhesions in living cells in several different contexts.   Figure ...substrate furimazine. Right: Demonstration of PILATeS. Cell adhesion to a PILATeS-functionalized coverslip creates...

... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...amplitude measured for each droplet in each well (Figure 2). A clean ddPCR should have a clear separation between...as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has ...used in the detection of rare sequences and single cell analysis. It is also being used in the detection...

...represents the time and intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength...most people use that setting. Figure 2: As the cell passes through the laser beam and causes it...proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line one ...Many cell types can be found in the tumor microenvironment (tumor cells, immune cells, blood cells and ...about 53% of my cells are dendritic cells and ~13% are CD8+ T cells. Around 34% of the cells are neither ...neither of the two cell types (likely B cells and others). Almost no cells stain positively for both markers...measure your cells of interest. In many experiments you’ll be working with a heterogeneous cell population...

...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser...ability to establish and maintain itself within host cells.  The plasmid has multiple ori that are functionally...eutrophus, Bartonella bacilliformis, Bordetella spp., Brucella spp., Caulobacter crescentus, Escherichia coli...

...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup...scFvs and T cell receptors and are expressed on T cells to make CAR T cells. The extracellular scFv portion...Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific antigen, it brings T cells to a ...recognizes an antigen of interest while the intracellular T cell receptor portion of the CAR facilitates ...scFv part of the receptor can activate T cells to kill cancer cells. scFv’s small size also allows for their...

... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...Zooming Forward Design: We produced a video celebrating Massachusetts Nonprofit Day, which was the perfect opportunity...  Quintin Marcelino...

...immunosurveillance." Cancer cell 30.1 (2016): 147-160. PubMed PMID: 27411589. 2. Todoric, Jelena, Laura Antonucci...represent a favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive...cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer, Inflammation and Immunity and...recent findings in basic immunology, cancer-immune cell interactions, cancer immunotherapy, as well as new...immunosurveillance. The presented study analysed a model of hepatocellular carcinoma, a cancer which can develop from ...example the upregulation of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment...follow the interactions between tumor and immune cells in real time. Addgene depositor Professor Max Krummel...

... variation (CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) ...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius... to date for visualizing genomic loci in living cells. Previous iterations of genomic loci visualization...labeling system to visualize chromosomal loci in human cells (Ma et al., 2015). To do so, they fused a different...visualization of sites on different chromosomes in living cells, the system requires high-copy chromosome-specific... aptamer system to visualize the gRNA in living cells. Broccoli aptamers are nucleic acid sequences that...production needed to deliver CRISPR-Sirius into cells. They named this guide sgRNA-Sirius-8XMS2 which ...

...otherwise be difficult to separate out. In Figure 2, you can see that, without the FMO, it would be difficult... from the Ly6C positive population. Figure 2: An FMO gating example. The FMO sample (A) can be ...and beads that are unable to bind to them: like cells but with far more predictability. The beads create...The benefit of beads is that they do not require cells from your experiment, and they are non-specific....flow cytometer, you generally want your positive cells to read between 104 and 105 in order to have enough...the plot to see all of your positive and negative cells. This can be achieved by adjusting your voltage ...gating schematic to easily determine where your cells of interest are (B).   Using FMO controls In...

... you can read more about them in review articles (2–4). You can also take a look at my recent paper to...Development 118, 401–15 (1993). PubMed PMID: 8223268. 2. J. B. Duffy, GAL4 System in Drosophila : A Fly Geneticist... to express one gene in one cell type and another gene in a different cell type (6–8). Although it’s often...systematically study gene function with temporal control and cell-type specificity! The Gal4/UAS system At a more...the hedgehog gene can be overexpressed in several cell types by using different Gal4 drivers. It is much...much easier to mate flies to get the right cell type expression as opposed to making a new fly with hedgehog... Targeted gene expression as a means of altering cell fates and generating dominant phenotypes., Development...

...is always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid...the presence of oxidizing agents. Adapted from Fig. 2 Campbell et al, Nature Methods   Find YFP plasmids...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...

... you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...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...

...mutation is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new prenatal test for sickle-cell anemia." ...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically advanced... of the mutated β globin gene that causes sickle cell anemia. The mutation happens to destroy a sequence...conducted on a sample from an individual without sickle cell anemia, two DNA fragments are observed because the... site where MstII cuts. In a patient with sickle cell anemia the enzyme does not cut the DNA so only one...