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  1. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...overlapping 5′ and 3′ EGFP fragments under the ubiquitous CAG promoter. The approximately 500 bp genomic fragment...cassette. (b) pCAG-EGxxFP target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target...cloned directionally into the BbsI site. (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330...Once amplified, you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques...of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with the individual gRNAs in...Remember the primers you designed to generate your pCAG-EGXXFP plasmid? They are the perfect primer sets...
  2. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...integration is TA dinucleotides and it leaves behind the CAG DNA footprint from its terminal sequences at the ...
  3. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...that AAV-PHP.eB, when paired with the ubiquitous CAG promoter, is relatively biased toward L5 and inhibitory...is quite pronounced, even when using a ubiquitous CAG promoter. Spatial transcriptomics shows that its ...
  4. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...Autobow systems are available with either the Thy1 or CAG promoter, while Brainbow AAV is under control of ...target sequence in between two fragments of EGFP in pCAG-EGxxFP. The EGFP fragments contain 482bp of overlapping...
  5. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...mice. The Jacks Lab used this backbone to target a CAG-driven loxP-stop-loxP cassette into the Rosa26 locus...pCS2TAL3-RR Pawel Pelczar pCAG-T7-TALEN(Sangamo)-Destination series, pCAG-Golden-Gate-Esp3I-Destination...pCAG-Golden-Gate-Esp3I-Destination Takashi Yamamoto pcDNA-TAL-NC2, pCAGGS-TAL-NC2 Charles Gersbach pcDNA3.1-GoldenGate, pcDNA3.1...
  6. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...oligonucleotides Cas9, photocaged-gRNA Gene editing Manna et al., 2019   NPPOC-caged trimethoprim (TMP) ...transcription Zhou et al., 2020   NPOM-caged nucleobases Cas9, photocaged-gRNA Gene editing For more details...chromatin, and study kinetics. Using chemistry to photocage CRISPRs The aforementioned techniques each employed...These researchers used a genetically encoded photocaging technique to insert a light-removable protecting...protecting group, specifically a nitrobenzyl photocaged lysine (PCK), on the Cas9 protein (Hemphill et al.,..., read this article). The group first tested photocaging various lysines in Cas9 to determine which best... ability to cleave targeted DNA, settling on photocaging the K866 lysine, as seen in the figure below....
  7. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ...final Forward Primer sequence of 5'-TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design...final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA-3’. Preparing the PCR product for...
  8. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...adaptations of this method have been to tag, cage, uncage, and trap the chemical fluor with the genetically...
  9. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...localization sequence) and expressing it from a CAGGs promoter. However FLPe is still less efficient than...binding domain (LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These Cre/ FLP versions are retained...
  10. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...enzymatic activity. In addition, proteins can be “caged” by fusion to specific domains thereby rendering... The authors use PhoCl in combination with this caging technique to beautifully demonstrate light-dependent...
  11. #Deckthelab Contest Winners 2017

    Type
    Blog Post
    Published
    Jan. 31, 2018, 2:44 p.m.
    ...are some of our 2017 contest winners:   Chicagoland's Community Biology Lab (@ChiTownBio) and one ...
  12. Addgene's Website Update

    Type
    Blog Post
    Published
    April 8, 2024, 2:15 p.m.
    ...informed by Web Content Accessibility Guidelines (WCAG).  But don’t take our word for it — check out all...
Showing: 21 - 40 of 51 results