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We narrowed to 772 results for: INA

Showing: 51 - 60 of 772 results

  1. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...machinery making an original C-G pair converted to T-A or reverted to the original C-G by base excision...invented CRISPR-base editors by tethering cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick...indel formation. After binding with DNA, the CBE deaminates a target cytosine (C) into a uracil (U) base...., there are no naturally known 'DNA adenosine deaminases' which would theoretically give rise to an adenine...resulted in the successful evolution of an adenosine deaminase that can act on DNA for adenine base editing. ...created a mutant library of E. coli tRNA adenosine deaminases (ecTadA), fused it with dCas9, and transformed....          The artificially evolved adenosine deaminase catalyzes the conversion of a target ‘A’ to ‘I...

  2. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...site-specific serine recombinase. First, the prime editor installs the recombinase “landing sites,” then... MagIC-Cryo-EM for enrichment and structure determination LEA proteins protect samples during cryo-EM ...editing CHARM: A compact epigenetic silencer Recombinant antibodies for marking astrocytes MagIC-Cryo-EM...MagIC-Cryo-EM for sample enrichment and structure determination By Mike Lacy Conventional cryogenic electron ...MagIC-cryo-EM for sample enrichment and structure determination. A) Schematic of workflow. B) Detail of spacer...Funabiki, H. (2024) MagIC-Cryo-EM: Structural determination on magnetic beads for scarce macromolecules ...then the recombinase handles the installation of the donor sequence. The latest work evolves and engineers...

  3. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...phosphoserine into recombinant proteins, such as the kinase MEK1 (5). MEK1 is inactive without requisite...candidate regulatory kinase is known, it’s possible to generate non-phosphorylated recombinant protein and perform...in vitro kinase reactions. Often, however, researchers lack knowledge of the regulatory kinase, access ...access to active kinase for the reaction, or have difficulties with kinase specificity in vitro. The establishment...establishment of a system in which purified, recombinant protein could be produced with site-specific incorporation...incorporation of phosphoserine directly into recombinant proteins (5). Optimization of this system defined...create a pSer/phosphomimetic dual mutant) elevated kinase activity over 70-fold over dual phosphomimetic ...

  4. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...first segments of the spinal cord showing the cortico spinal tract, CST, spinal cord and cranial nerve...Cuneate tract, dCST= dorsal cortico spinal tract, vCST= ventral cortico spinal tract, dc= decussation of the...was first pioneered by Voie and colleagues and originally named orthogonal-plane fluorescence optical sectioning...Spelman focused a laser beam into a thin sheet to illuminate a fluorescent sample and captured the reflected... lens oriented perpendicular to the plane of illumination (i.e. light sheet).  This is in contrast to ...described the single-plane or selective-plane illumination microscope (SPIM). In order to improve axial...optical sectioning of a specimen is based on discriminating the out-of-focus reflected light by using a...

  5. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...sites are eliminated by digestion/ligation and do not appear in the final construct. The destination vector...Type IIS enzyme and ligase. Although the original destination vector + insert may spontaneously religate...ligated following a heat inactivation step. If your genes of interest or destination vector contain multiple...correctly, the recognition sites do not appear in the final construct, allowing for precise, scarless cloning...complementary overhangs that direct assembly of the final ligation product. As shown below, a fragment with...overhangs. Entry DNA overhangs may be present in the original plasmid (Option 1) or added using PCR-based amplification...ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a single...

  6. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ... or under certain stimuli. Cre recombinase catalyzes the recombination of DNA sequences located between.../c, and 129. Conversely, outbred strains were originally derived from random crosses between mice of uncharacterized...Outbred strains of mice are useful for comparing dominant and recessive traits and their impact on diseases...or into embryonic stem cells. Non-homologous recombination incorporates this DNA randomly into the mouse...knockin strains are generated via homologous recombination. A targeting vector is electroporated into embryonic... ends to the breakpoint promotes homologous recombination of the targeting vector. CRISPR/Cas9-based systems... of small molecules that promote homologous recombination or the use of long single-stranded DNA have ...

  7. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...intrinsic bacterial recombination pathway, referred to as recA-independent recombination, joins together ...modifications requires multiple recombination events to assemble the final plasmid. At least 5 fragments...assembly can be achieved in vivo using a bacterial recombination pathway that is present even in common lab cloning... The pathway is ubiquitous, with successful recombination reported in all laboratory E. coli strains tested...Garcia-Nafria et al., 2016) uses the bacterial recombination pathway to allow any cloning procedure to be...strain: when asking the cell to perform multiple recombination events, all linear fragments need to get into...For simple cloning requiring just one or two recombination events, homemade competent cells of 107 CFU/...

  8. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...ligated into Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt...Originally published Jan 28, 2016 and last updated Sep 10, 2020 by Jennifer Tsang. CRISPR makes it easy...ordered assembly of multiple components into a destination vector. The first step in CRISPR/Cas9 Golden ...together to allow for ordered assembly into a destination vector that expresses Cas9. This is illustrated...drive ordered assembly into a Cas9-containing destination vector. Note: depending on which method you use...fewer than four gRNAs, you’ll clone in a polyT-termination sequence for each unused promoter. This step ...to generate all of the overhangs needed for the final ligation step. Plasmids are then digested using ...

  9. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...delivery By Cythina Arokiaraj (Gradinaru lab) Another useful viral tool deposited by the Gradinaru lab, AAV1...recommends these recombinant antibodies for use in western blots, though the original parent clone (M2...primate brains Endothelial-specific AAV delivery Recombinant anti-FLAG tag antibodies Peptide-assisted genome...mitochondrial membranes imaged with structured illumination microscopy in HeLa cells expressing COX8a-mStayGold... photostability and brightness compared to the original, though its maturation is even faster. Published...maintaining the bright and stable fluorescence of the original (Ivorra-Molla et al., 2023). They also named their...Hridaya Patel (Addgene co-op) Researchers in the Gradinaru lab have developed an engineered adeno-associated...

  10. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    Published
    July 16, 2020, 1:15 p.m.
    ...though the data is heavily dominated by the U.S. (42%), Europe (23%), and China (14%). Spikes in plasmid...instabilities in the China data, so we recommend interpreting the model results for China with more uncertainty...entered lockdown for weeks to months, pausing or terminating employment for a significant fraction of the ...encountered COVID-19 at different times. Most notably, China was affected by COVID-19 earlier than the rest of...hierarchical model: Because the U.S., Europe, and China account for over 75% of all of our plasmid orders...COVID-19 event windows as 01/15/20 -- 03/15/20 for China and 03/11/20 -- 05/19/20 (the last date in our dataset...day across regions, with Europe most impacted and China least impacted. The inferred weekend effect is, ...
Showing: 51 - 60 of 772 results