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We narrowed to 180 results for: ENDO 2

Showing: 81 - 100 of 180 results
  1. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids... Targeted DNA demethylation and activation of endogenous genes using programmable TALE-TET1 fusion proteins...-specific editing of histone modifications at endogenous enhancers. Nature Biotechnology 31:1133–1136 ...
  2. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...base pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles that...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage...
  3. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...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...produce a similar product, but do not target an endogenous gene in your experimental cells. For example,...
  4. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...GreenGate are type IIS restriction endonucleases. They are like other endonucleases except that they cut at arbitrary...be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very sensitive... They finally traced the problem back to the endonuclease. “It’s just a sensitive protein,” he says. “...
  5. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    Published
    March 8, 2021, 2:41 p.m.
    ... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of... to the controls listed on the vendor’s website. Typically, the vendor will demonstrate that their antibody...efficiently binds to endogenously expressed protein at physiologically relevant levels. Some vendors will also test...targeting your protein of interest from a variety of vendors and you may even find plasmids encoding antibodies...attention to the validated applications listed on the vendor’s website and choose an antibody that has been validated...antibody binds to protein of the correct size. Many vendors will show that the antibody binds to transiently...
  6. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...OptoTIAM (magenta) expressed in a monolayer of endothelial cells, showing cell protrusions and overlaps ...junction integrity in the vascular barrier of the endothelium, but they could be applied to study other actin-dependent...provided CIB constructs for targeting Golgi, ER, and endosome membranes) (Figure 4). The authors used SuperPLD...scale, enabling precise control over vascular endothelial barrier strength. eLife, 12, RP84364. https:/...
  7. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...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 ...lab has the capability) with receptor mediated endocytosis. They used this to edited those brown marmorated...
  8. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...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...Physical and genetic studies with restriction endonucleases on the broad host-range plasmid RK2. Molecular...
  9. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...conjugated to molecular species with high affinity for endogenous molecular targets such as paclitaxel. Unlike ...required as the small-molecule binding motif targets endogenous protein (Figure 1f). Labeling strategy pros...
  10. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...typically expressed at much higher levels than endogenous proteins. Data obtained from experiments using...tagged protein may not be translatable to the endogenous system and will need to be verified using alternative...
  11. Hot Plasmids - January 2023

    Type
    Blog Post
    Published
    Jan. 31, 2023, 2:15 p.m.
    ...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...also demonstrated the ability to be used as an endogenous RNA imaging tool.   Fig. 1: Csm, shRNA...
  12. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ... as muscular dystrophies, heart disease, and type 2 diabetes, across four different types of human cells...(#186715) and hCtRNA_VT (#186716).   Fig. 2: Comparison of previous representative dual-gRNA architectures...expression, the individual gRNAs are released by endogenous tRNA processing machineries, specifically, RNaseZ...
  13. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   ...
  14. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ... is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...biology98.3 (1975): 503-517. PubMed PMID: 1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new...cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae." Proceedings of the...
  15. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R ...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular... of certain expressed proteins endA, endA1 Endonuclease I (nonspecific cleavage of dsDNA ) mutation ...
  16. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...PMID: 24086725. PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher...recommends using an anti-mCherry antibody rather than endogenous mCherry fluorescence when visualizing hM4D(Gi...
  17. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...it as “a novel mode of optogenetic control of endogenous cellular processes." The LITE system can now ...Konermann et al. "Optical control of mammalian endogenous transcription and epigenetic states." Nature....
  18. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...this method, but your edit doesn’t affect any endogenous restriction enzyme sites? Never fear! You can...Resources References Eva K. Brinkman, Tao Chen, Mario Amendola, Bas van Steensel. Easy quantitative assessment...
  19. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    Published
    Feb. 11, 2021, 2:15 p.m.
    ...plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 gamma subunits of the heterotrimeric G...activation via bioluminescence resonance energy transfer 2 (BRET2). Check out our blog post on GPCR signaling...chemogenetics as they are relatively insensitive to their endogenous ligand, acetylcholine, and are only activated...
Showing: 81 - 100 of 180 results