...loci targeted One Many Sample type(s) Cell lines Cell lines, primary cells Table 1: Key differences...multiple targets because different CAPTURE cell lines will have varying levels of dCas9 and gRNA expression...components eliminates the need to create stable cell lines and allows CAPTURE 2.0 to be used with primary cells...like to interact with, and how many promoters or genes each enhancer is interacting with. Super enhancers...

...SB_loop, SB_convex. These scaffolds contain serines and threonines at positions that can be randomized for...epitopes that are hard for antibodies to get to, like ones that are hidden in the molecular crevices proteins...Developing antibodies against proteins embedded in membranes is tricky because they have fewer hydrophilic ...diverse pool of sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger...

... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...changes under magnetic field. Such changes could be harnessed or enhanced for future MagLOV-based tools, just... Ingaramo, M. (2024). Magnetic control of the brightness of fluorescent proteins. Zenodo. https://doi....., & Kusamoto, T. (2023) Single-Molecule Magnetoluminescence from a Spatially Confined Persistent Diradical...

...Origins of protein tags For about as long as the disciplines have existed, cell and molecular biologists have...developing generic strategies for detecting cloned genes (Munro and Pelham, 1984). The strategy they landed...1985). Myc was one of the earliest identified oncogenes and was a target of intense research. Evan, et...explore Myc’s role in cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding...tagging to detect proteins expressed from cloned genes: deletion mapping functional domains of Drosophila...

...a chance of seeing a phenotype since additional genes that can do the same function are less likely to...Essentially, the X, second, and third chromosomes are the ones you’ll work with. If YFG is somehow on the Y or ...Geneticists need a method for maintaining stable lines of genetic mutants, including in situations where...lose track of your mutation. To maintain stable lines of flies carrying a specific mutation, Drosophila...Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective. Their Snapdragon tool helps...

...contacts that bring apposing membranes together and can wrap the membranes into stacked structures including...every lab in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to...tubules can be grossly distorted into dense stacked membranes termed Organized Smooth Endoplasmic Reticulum ..., M.V., et al., Fluorescent proteins from nonbioluminescent Anthozoa species. Nat Biotechnol, 1999. 17...

...” is the title of the chapter that examines the inherent brokenness of scientific publishing and research...tackles much broader issues in this brief volume. He examines, amongst other topics: racism and sexism in science...the U.S. environment, and a decline in the competitiveness of U.S. science and technology.” This last ...in under 200 pages is an impressive task to set oneself. As a scientist who enjoys learning a little bit...

...(1x cysteine) pTP955 (3x cysteines) Mouse IgG1 TP886 pTP943 (3x cysteines) Mouse IgG1 TP1107 pTP1112...1x cysteine) TP1122 (3x cysteines) Mouse IgG2a TP1129 pTP1005 (3x cysteines) Mouse Kappa TP1170 ... pTP1174 (1x cysteine) pTP1172 (3x cysteines) *Rabbits have only one IgG subclass.   Express the ...

...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...action of the most common antibiotic resistance genes and which applications are best (or worst) for each...plasmid of interest, preventing this issue. The genes and their antibiotics   Ampicillin Ampicillin, often...antibiotic due to stable integration of resistance genes in their genomes. If you need a plasmid that can... Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage...

...breast and ovarian lines commonly used in lab. It is recommended you check your cell lines for HR-related ...covered in this blog, with tips and tricks for harnessing the homology-directed repair pathway, designing...surprise then that these edits take more planning and finesse then their knock-out counterparts. To kick off ...experiment today! Get ready to identify knock-in clones by reading up on validating genome edits to find...

...biological disciplines and scientists employ GFP for a wide number of functions, including: tagging genes for...Bioluminescence and fluorescence from proteins such as Green Fluorescent Protein (GFP) has likely existed...began engineering new versions of GFP through mutagenesis in order to improve its physical and biochemical...Click here to view Addgene’s collection of empty backbones for constructing fluorescent fusions. Transcription...

... propagation of themselves and their associated genes. These systems better ensure that plasmids are passed...death. The team used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed... (hok), a long-living toxin that damages cell membranes; Suppression of killing (sok), a short-lived RNA...proof of concept, they tested a lux-mediated bioluminescence reporter in vivo in the absence of selective... Ogura, Teru, and Sota Hiraga. "Mini-F plasmid genes that couple host cell division to plasmid proliferation...

...To deliver genes using lentiviral vectors, you need an envelope protein on the virus’s surface and a ...stomatitis virus, HIV-2, and human foamy virus in cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal... were less stable than viruses produced in cell lines that do not express these sugars. (Takeuchi et al...study the spike protein’s role in infecting cell lines without using intact SARS-CoV-2 virus. Scientists...find tables and information on different viral membranes, tropism, and advantages and disadvantages: Table...

... similar brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP...Master. Note: BRET= Bioluminescence Resonance Energy Transfer CRET= Chemiluminescence Resonance Energy Transfer...that can affect FRET measurements include: the brightness of a fluorophore pair, donor:acceptor stoichiometry...

... are essential for processes such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction...reactive and forms a link with nearby nucleic acids (McInnes et al., 1990). Photobiotinylation is sometimes ...18988027. PubMed Central PMCID: PMC3223083. 6. McInnes, J. L., Forster, A. C., Skingle, D. C., & Symons...Biotinylation ThermoFisher Scientific: Biotinylation Guidelines  ...

...Isolation and Concentration (MagIC)-cryo-EM that combines enrichment and imaging of proteins captured on...with a GFP nanobody (Figure 1). This enrichment streamlines the purification and minimizes sample loss while...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that... to partially correct mouse models of genetic blindness (An et al., 2024) — the first demonstrated use...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...

...Repair is Used to Edit Genes Read How Non-Homologous End Joining is Used to Delete Genes Additional Resources...exons), but much of the sequence appears to be nonessential. Within the mutational hotspot for dystrophin... cells expressed functional dystrophin. Editing genes in vivo is of course much more difficult than in...these different mutations. For each therapy, effectiveness and off-target risk must be evaluated separately...

...plasmid yield and/or DNA quality. Table 1 below outlines a few of the more common genetic changes found...encodes proteins needed for bacterial conjugation. Genes listed on F´ are wild-type unless indicated otherwise...lac Lac operon mutations Blue/white screening of clones   lacIq lac repressor overproduced, expression...transformation efficiency deoR constitutive expression of genes for deoxyribose synthesis Allows uptake of large...)]  Note: Inactivating mutations in specific genes are signified with a minus sign (-) as is typically...

...edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post...protein-coding capacity of a locus. When using S. pyogenes Cas9 (SpCas9), potential target sites are both...Veres et al., 2014). When working with single-cell clones, the authors note that “clonal heterogeneity may...obstacle to the generation of truly isogenic cell lines than nuclease-mediated off-target effects.” Further... CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing. Cell Stem ...

...Finally, as Pamela Hines of AAAS points out on the panel, irreproducibility undermines public trust in science...Last Wednesday we worked with the Harvard GSAS Science Policy Group to organize a Minisymposium on Reproducibility...) - Panel Alex Tucker (Ginkgo Bioworks) Pamela Hines (Senior Editor at Science) Edward J. Hall (Professor...journalists and scientific experts discussed by Pamela Hines Minisymposium on Reproducibility Booklet - Including...