We narrowed to 813 results for: cell
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TypeCollection...collection of synthetic biology plasmids related to cell-to-cell communications and environmental sensing. Sensing...
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What's New in CRISPR - Spring 2019
TypeBlog Post...temperature incompatible to genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus...can block Cas12a-mediated genome editing in human cells. Read the paper in Science Find the anti-CRISPR...anti-CRISPR expression plasmids CRISPR-based method for cell lineage tracing Valcyrie is a CRISPR-based marker...marker excision method for marking distinct cell clones developed by the Michalis Averof lab. It’s an alternative... -
Hot Plasmids: Winter 2025
TypeBlog Post... 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..., CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only...recordings of spontaneous activity in human stem cell-derived neurons expressing ASAP5-Kv. C) Cultured... -
MXS Chaining
TypeBlog Post...construct was then introduced to HeLa cells (Figure 1). The resulting HeLa cells show robust labeling, with strong...protein localization, as well as cellular processes like gene expression and cell migration (Sladitschek and... MXS-chaining were used to label subcellular structures in live cells. Four individual expression constructs...imaging or flow cytometry applications in mammalian cells. The modules used in this method include fluorescent...components, one can generate constructs to visualize cell cycle dynamics, titrate inducible transgene expression...applications in flow cytometry approaches in mammalian cell culture systems (their original goal). Their work...detection of each fluorophore at the respective subcellular structure (Figure 1). Here, MXS Chaining gave... -
Tips for Using FRET in Your Experiments
TypeBlog Post...(FRET) microscopy imaging of live cell protein localizations. J Cell Biol. 2003 Mar 3;160(5):629-33. ...measurement of single molecule interactions in living cells using a microscope. Historically, molecular interactions...suitable for studying interactions within living cells. By coupling fluorophores to proteins, pioneers ...directly detect protein/protein interactions in living cells. Since then FRET has also been used to measure conformational...efficiency and cross-talk. Methods to Measure FRET for Cell Biology Studies Several methods have been used over...microscopy: monitoring protein interactions in living cells. Bioessays. 2012 May; 34(5):341-50. doi: 10.1002... Constructs to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian... -
RaPID Detection of RNA-protein Interactions
TypeBlog Post...proteins that bind an RNA motif of interest in living cells. Challenges with existing RNA-protein interaction...and naturally occurring complexes present in the cell, it can also introduce non-physiological RNA-protein...a short timescale, such as during drug action or cell signaling. Limitations of RaPID Like other BirA*... controls include both scrambled RNA motifs and cells transfected with BirA* protein alone (i.e. no RNA...protein that’s highly expressed in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels...2018). RNA–protein interaction detection in living cells. Nature Methods, 15, 207-212. PubMed PMID: 29400715... -
Viral Vectors 101: Virus Safety
TypeBlog Post...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:... the procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-...vector carries an shRNA to a gene essential for cellular function, this would also be a higher risk vector... -
Plasmids 101: Biotinylation
TypeBlog Post...chemically. Enzymatic biotinylation occurs naturally in cells and is the process by which biotin is covalently... Immunosorbent Assay) Southern/Northern blotting Cell surface labelling Attaching polypeptides to a substrate...substrate Flow cytometry/fluorescence-activated cell sorting (FACS) In situ hybridization Methods for...proteins in bacteria, yeast, insect and mammalian cells. Check out Kay et al., 2009 for methods on biotinylating...Application of avidin-biotin technology to improve cell adhesion on nanofibrous matrices.” Journal of Nanobiotechnology...purification of tagged transcription factors in mammalian cells and transgenic mice.” PNAS 100.13 (2003):7480-5....-H., Lee, C.-Y., & Li, P.-C. “Buoyancy-Activated Cell Sorting Using Targeted Biotinylated Albumin Microbubbles... -
"Build Your Plasmid": The Game - Play to Teach
TypeBlog Post... of human stem cells when students are not even allowed to manipulate prokaryotic cells? Especially, if...where you need to apply basic molecular biology, cell biology, microbiology, genetic engineering and strategic...could be used to genetically manipulate human stem cells. Then they had 10 minutes to expose their strategy...construct and explain why they selected a certain target cell population. Who is the winner? Ultimately, society... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post... of all of the mRNA bound-ribosomes in a cell. When cellular mRNAs are digested with ribonucleases, ribosomes...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...imaging the first round of translation from single cells to living animals.” Science 347(6228) (2015): 1367...of Translation of Single mRNA Molecules In Vivo.” Cell 165(4) (2016): 976-89. PMID: 27153498 PMCID: PMC4889334...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... -
Optimizing Donor DNA for Enhanced CRISPR Genome Editing
TypeBlog Post... between Cas9 (which causes the breaks) and host cell DNA repair factors (which repair those breaks) makes...example, correcting the allele that causes sickle cell anemia. However, HDR occurs much less frequently...a structure could drive sequence replacement in cells. Targeting three dCas9 molecules to a precisely ...functional Cas9 protein and the targeting guide RNA into cells by electroporation. Results from our lab and others...Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins... -
Plasmids 101: Using Transposons in the Lab
TypeBlog Post... yeast, plant, insect, and mammalian cells, including human cells. piggyBac is biased to integrate at ...is active in vertebrates and integrates in human cells at rates similar to retroviral vectors. The hyperactive...piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive PB...transposase (hyPB) is ~10-fold more active in mammalian cells compared to the codon-optimized wild-type piggyBac...G/C)TNA. Tol2 can deliver 10-11 kB to mammalian cells without decreased efficiency, with a maximum cargo...and has lower efficiency of integration in human cells than piggyBac and Sleeping Beauty. Minimal Tol2 ... or barcodes. When transposons are delivered to cells or a model organism, they insert into the host’s... -
Plasmids 101: Modular Cloning Applications and Kits
TypeBlog Post...can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants...transport studies in HEK cells, recombinant protein purification in bacterial cells, and a two-hybrid protein...for synthetic biology applications in bacteria or cell-free systems. The approach is particularly useful...add-on kit and is compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo ...of expression and integration needed in mammalian cell types, and comes with a large library of common ...protein interaction study in yeast cells. Need more MoClo? Check out Addgene's MoClo Guide! Customizing...Construction of Biological Circuits in Mammalian Cells.” ACS Synthetic Biology, vol. 8, no. 11, American... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post... 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... -
Educational Planarian Tools for Teaching Developmental Biology Techniques
TypeBlog Post...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...specific cell types or entire organs in planarians. For example, the gene Piwi labels neoblast cells and the...techniques to alter gene expression or to label specific cells, but there are too few resources to teach students... in tissue. This is useful for labeling specific cell types and for viewing how gene expression changes... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog Post...circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable for up to...possible and submitted our manuscript to the journal Cell just six weeks later. The Zika publication timeline...of Toronto. His research is focused on developing cell-free synthetic biology applications for human health...Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from...de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939. PubMed PMID: 25417166. PubMed...Keith, et al. "Paper-based synthetic gene networks." Cell 159.4 (2014): 940-954. PubMed PMID: 25417167. PubMed... -
Adenovirus Plasmids
TypeCollection...target a broad range of dividing and non-dividing cell types with almost 100% efficiency. Unlike lentiviruses...recombination of adenovirus plasmids in either mammalian cells or microorganisms, including bacteria and yeast....can then be transfected into mammalian packaging cell lines to generate adenovirus particles. Read our... -
p53 Pathway
TypeCollection...p53 are in promoting cell cycle arrest, apoptosis, or senescence in damaged cells. The p53 name arose ...or E2 Cyclin G CCNG1 CCNG2 Cyclin G1 or G2 CDC25 Cell division cycle 25C Cdk4/6 CDK4 CDK6 Cyclin-dependent...member 10b E2F-1 E2F transcription factor 1 Fas Fas cell surface death receptor Gadd45 GADD45A GADD45B GADD45G...transcription factor and tumor suppressor activated by cellular stress. Cancer...transcription factor and tumor suppressor activated by cellular stresses like DNA damage, oxidative stress, and... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog Post...Syndrome and Related Disorders. Cell 152:984–996 . https://doi.org/10.1016/j.cell.2013.01.038 Guy J, Gan J, ...deliver the precise amount of protein to each brain cell to restore physiological levels. Much research is...repair a gene in a living animal across multiple cell types using a mouse model for a human neurological... that enough wild-type MECP2 is made inside the cells, but this is exciting progress in tackling the root...Resource Center. Researchers can also find links to cell line repositories including patient-derived iPSCs...Modifications in Human-Induced Pluripotent Stem Cells. Front Genet 10: . https://doi.org/10.3389/fgene...Neurological Disorder by Programmable RNA Editing. Cell Reports 32:107878 . https://doi.org/10.1016/j.celrep... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog Post...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...Hawaiian bobtail squid has an extreme number of nerve cells and an unusually high level of RNA recoding in the...using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all (maybe because...total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little bits of hydra can...Mormyridae (Jason Gallant) Weakly electric fish are excellent systems to study for convergent evolution because...