...product, the transformation may not result in any cells, or all the colonies screened might not contain ...verify the strain and the efficacy of your competent cells (when they get old, they lose competency). If all... look at your experimental design. Full lawn of cells. The antibiotic in your plate might not work. Check...acknowledged in hundreds of publications including Cell, Nature, and other journals.   Paolo Colombi is ...

...interaction networks of specific proteins in living cells. Until recently, the enzymes most frequently used...but requires H2O2, which can be toxic in living cells. BioID was derived from biotin ligase and uses far...Alexander Green lab found a solution: a low-cost, cell-free detection assay that can be completed without...family, known to be involved in cytoskeletal control, cell growth, and endocytosis. Recently, the Sidhu group...to studying membranes, organelles, and other subcellular compartments is via proteomics--the study of ...

...also gives cells antibiotic resistance to neomycin so that you can select the population of cells with the...Only a small percentage of the proteins inside a cell have a commercial antibody against them. There’s...antibody. It’s an epitope not found in human or other cells. It was made artificially. That means you aren’t...

...successfully infected/expressing cells by flow cytometry. The isolated cells were injected into mice and imaged...to changes in intracellular calcium and is useful for direct imaging of events in cells and brain explants...packaged into a retrovirus for infection of host cells. The fluorescent moiety of the BRET reporter was...imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs) can be problematic...available through Addgene (Plasmid #83926). Live-Cell Voltage Sensor  Ingaki and colleagues (8) were interested...including monitoring multiple cellular events such as the dynamics of subcellular structures and gene expression...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors...

...to enable three-feature cell targeting. C) Example of viral targeting of cells expressing Cre AND Flp,...parameters. Figure 1: Examples of intersectional cell population definitions using recombinases. A) Cre...adeno-associated virus (AAV)-borne payloads to be expressed in cells based on a doubly-specified combination of genetic...Neve R, Huang ZJ, Deisseroth K. 2014. Targeting cells with single vectors using multiple-feature Boolean...Cortex Reveals Local and Long-Range Circuit Motifs. Cell Rep. 28(13):3450-3461. PubMed (Link opens in a new...well-defined cellular sub-populations has generally been limited to single-component cellular definitions...multiple recombinase-expression strategies to define cellular sub-populations of interest, including dual-transgenic...

...Zhang Lab Plasmids The Human TFome Library Stem Cells Protocols & References MORF Plasmids MORF Pooled...Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F. Cell 2023 Jan 5; 186(1):209-229. PubMed (Link opens in...be used for screening transcription factors for cellular programming or cancer drug resistance. Depositor...transcription factors for applications such as cellular programming or cancer drug resistance. Protocols...

...function using transcriptional recording sentinel cells! 🎵 On the 11th day of CRISPR, Addgene gave to me...activation and interference screens for primary human T cells🎵 On the 3rd day of CRISPR, Addgene gave to me: ...Enhanced prime editing systems by manipulating cellular determinants of editing outcomes🎵 On the 5th ...

...Lentivirus can transduce both dividing and non-dividing cells without a significant immune response. These viruses... in the viral particle) is delivered to a target cell upon infection. When lentivirus is used for research...the researcher wants delivered to specific target cells. This genome is encoded by plasmids called "transfer...encoded by the transfer plasmid is toxic to mammalian cells, it can be difficult to efficiently produce viral...

...Worms are particularly well suited for studies in cell biology, development, and aging. Scientists can .... elegans hermaphrodite consists of 959 somatic cells, which makes them attractive for imaging and fluorescence...researchers to express desired constructs in precise cell types and/or developmental timeframes. Here we highlight...

...e.g., pHelper) and specific packaging cell lines (e.g., HEK293 cells) during AAV production. Read our AAV...AAV can transduce both dividing and non-dividing cells with a low immune response and low toxicity. Although...

...repair fix DNA double-strand breaks?  In eukaryotic cells, repair of DSBs occurs primarily by two pathways...used, however, these are generally more toxic to cells than their plasmid counterparts. Another consideration...how to fix a broken relationship. Pardo B, et al. Cell Mol Life Sci (2009). PubMed. Regulation of DNA...break repair pathway choice. Shrivastav M, et al Cell Research (2008). PubMed. Mechanisms of DNA double-strand...Improve HDR Efficiency for CRISPR Editing in Human Cells Resources on Addgene.org Browse All CRISPR Plasmids...double-strand break (DSB). DSBs can be caused by intracellular factors such as nucleases and reactive oxygen...

...Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Find plasmids...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...

...and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org... like proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et...epitope and transcriptome measurement in single cells. Nat Methods 14:865–868. https://doi.org/10.1038...

...slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing).  Some...enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol 33, 985–989 (2015). https://doi....Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021). https:/...deactivation with photocleavable gRNAs. Molecular Cell 81, 7, 1553-1565 (2021). https://doi.org/10.1016...evolved ways to prevent degradation of important cellular RNA molecules with post translational modifications...transfection efficiency and even track localization in cellular experiments. There are many suitable dyes to choose...

...delivered using AAV injection, the majority of the cells surrounding the injection site will become infected... ensure that the reporter remains silent until a cell or tissue-specific Cre is provided. Since the FLEx...used to study the physiology or behavior of certain cell populations.  Acknowledgement:Through a partnership...FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping...for monitoring Cre-mediated recombination at the cellular level in the mouse. 2003 PubMed PMID: 12665802...

... be expressed in the same cell, but not necessarily at the same subcellular compartment as the opsin or...modulate the activity of neurons and other brain cells. Since the opsins or chemogenetic receptors used...neuropil conceals identification of opsin-expressing cells). For other things to keep in mind when making fluorescent...cerevisiae by use of an epitope addition method. Mol Cell Biol 8:2159–2165 . https://doi.org/10.1128/mcb.8.5.2159...

...in transfected eukaryotic cells, allowing for them to be used as intracellular inhibitors.  Aptamers are...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 ...

...plasmids to upregulate their expression in specific cells. With a more concrete understanding of the molecular...molecules targeting SMN1 and its partners in mouse cell lines. Dr. Di Schiavi hopes that these studies will... of ~30 patient derived peritoneal mesothelioma cells lines and assess their tumorigenesis. These experiments...

...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 ...

... tools that effectively transduce many mammalian cell types. AAV is used in a wide range of experiments...Tetracycline Transactivators, Affinity Purification, Cell Ablation Monosynaptic Neuronal Tracing Rabies Virus-Based...

...Initiative Collection Cell Migration Consortium Center for Genome Engineering EXtracellular Plasmid RESource...Mammalian RNAi Microbiology Plant Expression Stem Cells Synthetic Biology Tet Inducible Expression Worm ...

...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 ...

...Can I directly transfect cloning grade DNA into cells? Cloning grade DNA is designed for use in cloning...cloning applications. Direct transfection into cells is not recommended. Can I order more than 2 µg of cloning...

... question about what should go into a cell, hover over the cell and instructions will pop up. When you...spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options for you to choose from, while... into the appropriate full and partial sequence cells. *Note: If you believe you need to deposit a collection...

...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...

...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...

...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,...

...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...