We narrowed to 819 results for: cell
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TypeCollection...and display of proteins on the extracellular surface of S. cerevisiae cells His-tagged versions of pFastBac... the elements needed for replication in the host cell and are designed for a user to insert a gene of ..., or express the insert they carry in the target cell. These vectors allow you to test the function of....1-mCherry - Expresses mCherry tag in mammalian cells pBI-MCS-EGFP - Tet-inducible Find more Tet-inducible...experiments, scientists often want to create stable cell lines in which the expression cassette of interest...is incorporated into the host DNA. For mammalian cells, you can do this through viral vector delivery...transduce both dividing and nondividing cells, integrate into host genome pLenti CMV GFP DEST ...
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Some Like it Hot: Thermostable GeoCas9
TypeBlog Post...mammalian cells. Harrington et al tested GeoCas9’s editing skills by transfecting HEK293 cells with GeoCas9...choice for a number of model organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs... 45 °C 50 - 70 °C Genome Editing in Mammalian Cells? Yes Yes Longevity in Serum?* No Yes Table ...experimental results. GeoCas9 RNP editing in mammalian cells Besides being able to take some heat, GeoCas9 can... -
Choosing Your Perfect Empty Backbone
TypeBlog Post..., or express the insert they carry in the target cell. These vectors allow you to test the function of...If you are planning to track your protein in the cell or purify it – and you do not have a specific antibody...selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent fusion ...get away with generating transiently transfected cells, you will not need a mammalian selection cassette.... Other vectors can be expressed in fly culture cells like S2 and carry the metallothionein promoter (... -
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... -
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... -
Caltech Systemic Capsids
TypeCollection...then cell type. For CNS: For infant rhesus macaques: Broad tropism - AAV.MaCPNS1, AAV.MaCPNS2 Cell type-biased...tropism - AAV.CAP-B22 Cell type-biased: Neurons - AAV.CAP-B10 Brain vascular cells - AAV.CAP-Mac For rats...tropism - AAV-PHP.eB Cell type-biased: Neurons - AAV.CAP-Mac Brain vascular cells - AAV9-X1.1, AAV1-X1...including: the delivery method, mouse strain, target cell type or tissue of interest, and model organism under... above helps you choose the best capsid for your cell type. First, do you want to target CNS ...Broad tropism - AAV-PHP.eB, AAV-PHP.AX, AAV-PHP.eC Cell type-biased: Astrocytes - AAV-PHP.AX with GRE or...AAV-PHP.eB with GRE or transgenic animal Brain vascular cells - AAV9-X1.1, AAV1-X1 Neurons - AAV.CAP-B10, AAV-... -
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... -
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... -
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... -
Antibody Production
TypeCollection...mammalian expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the.... This buffer is not compatible for use in live cells and will interfere with conjugation. For applications...antigen is first transiently expressed in mammalian cells. To learn which specific QC experiments were performed...email [email protected] . Immunocytochemistry (ICC) Cells are fixed and labeled with the antibody and a species-specific...Western Blotting (WB) Proteins are extracted from cells, separated by SDS-PAGE, transferred to a membrane... -
Antibody Plasmid Collection
TypeCollection...expressing these in bacteria, mammalian cells, yeast, plants, or insect cells. Benefits include consistency between... research purposes include animals or hybridoma cells, but are difficult to generate due to expense and...monoclonal antibodies (R-mAbs) expressed in mammalian cells. Monoclonal antibodies are derived from an individual...transiently transfected into mammalian tissue culture cells, the R-mAbs are secreted into the culture media ... -
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... -
"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... -
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... -
CRISPR Plasmids - Mammalian Expression
TypeCollection...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ... time and are potentially heritable in dividing cells. ID Plasmid Gene/Insert Selectable Marker PI Publication...DNA. Type VI enzymes that function in mammalian cells can be used to attentuate RNA levels. In mammalian...genomic loci using fluorescent microscopy in living cells. Design your gRNA sequence to direct the dCas9-FP...Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both... -
CRISPR Pooled gRNA Libraries
TypeCollection...experiment, target cells are treated with the pooled library to create a population of mutant cells that are then...quantities of cells and therefore, may not be suitable for use when the number of cells is limited (e....ribosomal, cell cycle) 51043 — 51048 Knockout Human Sabatini and Lander 3rd 10 Varies FNLCR CRISPRa Cell Surface...e.g. primary cells). Most CRISPR libraries require the use of lentivirus to deliver the gRNA/Cas9 to target... target cells. Therefore, you must be equipped and authorized to use lentivirus in your lab. Once you ...organism, and the library is only compatible with cells derived from that organism. All of our current pooled...140219 Knockout Human J. Chen 3rd 10 4,530 Murine Cell Surface Protein Knockout (McspKO) Library 200445...