Skip to main content

We narrowed to 223 results for: ENDO 2

Showing: 121 - 140 of 223 results
  1. Let There Be LITE Plasmids

    Type
    Blog Post
    ...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....
  2. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...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...
  3. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...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...
  4. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ..., et. al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...
  5. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ... strategy for linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...Science. 353, aad8559 (2016). PubMed PMID: 27463678. 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...
  6. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...without leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing...with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D...genome following donor removal, which alters the endogenous DNA sequence and has been suggested to cause ...
  7. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    ...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...stereotaxically, STX) to the brain. Figure 2: Selection, prep, and testing of AAV enhancers in ...Science (AIBS). Scientists at AIBS have allocated tremendous time and resources to generate highly specific...The Promoter Region – Let's Go! Plasmids for Endogenous Gene Tagging in Human Cells Resources on addgene.org...
  8. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...protocols that use antibody fragments to block the endogenous immunoglobulins and reduce the background staining...
  9. Components of CRISPR/Cas9

    Type
    Blog Post
    ...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...Cas9 genome editing systems use a non-specific endonuclease (Cas9 or closely related Cpf1) to cut the genome... have increased targeting specificity). Cas9 endonuclease functions While native CRISPR/Cas systems have...foreign DNA as well as the RNA guides required for endonuclease function, when used for genome engineering, ...the only CRISPR protein required is the Cas9 endonuclease or a variant thereof. This individual protein...) of a protospacer adjacent motif (PAM) Cas9 endonuclease binding to the target genomic locus is mediated...double-strand break (DSB) Cas9 and its variants have two endonuclease domains: the n-terminal RuvC-like nuclease domain...
  10. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...somewhere else (Fig. 2). Class II TEs have LTRs on both of their ends.    Figure 2: Overview of DNA...has four major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8, Cas7, Cas6) that forms...often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities.    ...
  11. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ... dCas9, a CRISPR protein variant lacking its endonuclease ability, to bind to genes without editing the...
  12. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...physiologically relevant in neurons, myocytes, and endocrine cells. It follows that the specific targeting ...
  13. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing... level observed will be dependent on both the endogenous promoter and the 3’ UTR of the locus of interest...
  14. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ... from E. coli is the presence of the protein endonuclease I encoded by the gene endA. Nowadays, the most...introduced in the purification protocol to remove endonucleases activity. For these reasons choosing the correct...
  15. Antibodies 101: Validation

    Type
    Blog Post
    ...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...that do not express your protein of interest endogenously and transfect them with a plasmid encoding your...ultimately planning to use your antibody to detect endogenous levels of the target in other cells or tissues...
  16. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach...detect the antigen of interest when expressed at endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches... since there can be large differences between endogenous expression and transfected overexpression, this...cannot confirm the antibody’s ability to detect endogenous levels of the target protein. It may only be ...overexpression. Additionally, if the cell line used has endogenous levels of expression, this can make the results...different antibody clones produced by different vendors. The specific epitope recognized by either antibody...
  17. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed monomeric fluorescent...
  18. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...antidepressants-like effects in rats." Psychoneuroendocrinology 46 (2014): 78-87. PubMed PMID: 24882160...
  19. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ... to: (1) prevent vendor lock in by making it easy to get data out of the ELN; and (2) facilitate inter-operability...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...
Showing: 121 - 140 of 223 results