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  1. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ... Although the lab did not conduct comprehensive genome-wide studies of ABE specificity, their other experiments... tightly constrained by PAM availability in the genome. So the development of near-PAMless Cas9 variants...significantly expands the library of accessible genomic targets for base editors (Walton et al., 2020)....2017). Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature, 551(7681), 464...Zhao, K. T., & Liu, D. R. (2017). Increasing the genome-targeting scope and precision of base editing with...2016). Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature... N., & Kleinstiver, B. P. (2020). Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9...
  2. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...alternative method that can be used for cloning or genome engineering and is based on homologous recombination...and will have more unintended changes to their genomes. 2) Designing ssDNA oligos that avoid MMR activation...when introduced into bacteria, integrates into the genome.  Mini-λ uses the endogenous red promoter pL and...stable integration of the phage into the E. coli genome. Selection and confirmation of recombinant clones...in Mammalian Cells CRISPR Methods for Bacterial Genome Engineering Resources on Addgene.org Browse our...
  3. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...integration of multiplexed constructs into target genomes and a 10-fold improvement in detection efficiency...efficiencies in a number of cell types at different genomic loci — in some cases, over 20-fold higher editing...abundant proteins and a critical component for viral genome packaging. Addgene recommends both for use in western.... mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy. Nat Commun., 14(1), 6578...
  4. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...generally applies to plasmids, but may also facilitate genome editing. Tailored mutations are commonly introduced...induced double-stranded break. This site-directed genome editing requires a template of high homology to...approach can be used to identify Cas9/CRISPR-induced genome modifications. Occasionally, multimerization of...Plasmids 101 Series Use CRISPR to Generate Specific Genomic Mutations Resources on Addgene.org Browse Our...
  5. Grad School Advice Part 2: Building Community

    Type
    Blog Post
    ... Ph.D candidate in the Program in Genetics and Genomics at Harvard Medical School. He studies developmental... a graduate of the PhD program in Genetics and Genomics at Harvard Medical School, and is currently a ...
  6. Grad School Advice Part 1: Picking a Lab and a Project

    Type
    Blog Post
    ... Ph.D candidate in the Program in Genetics and Genomics at Harvard Medical School. He studies developmental... a graduate of the PhD program in Genetics and Genomics at Harvard Medical School, and is currently a ...
  7. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...enzymes can knock down RNA without altering the genome, but Cas13s have higher on-target specificity. ... two guide RNAs to remove a given exon from the genome, and showed success in models of muscular dystrophy...CRISPR systems, only time will tell how large the genome and transcriptome engineering toolbox will be. ...characterized orthologs from “Anaerobic digester metagenome” (AdmCas13d) and Ruminococcus flavefaciens (nicknamed...
  8. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...at a specific genomic site, or randomly integrated (once or many times) into the genome. Make sure to ...lentivirus and retrovirus can package cDNAs for genomic delivery and have many therapeutic advantages, ...
  9. Let There Be LITE Plasmids

    Type
    Blog Post
    ...transcription-activator-like effectors (TALEs) as anchors for genome engineering applications was still very new. Now...role of specific epigenetic modifications in the genome,” Zhang said in MIT’s press release. In an accompanying...
  10. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...Interested in more genome engineering technologies? Browse others on Addgene's Genome Engineering Guide...genes, reporters, or other markers into a target genome. They did this by selecting the important elements...fusion proteins with a variety of genes across the genome. Although this tool was published in 2011, it was...scientists to use CRISPR-Cas technology to perform genome-wide knockout screens. These vectors and sgRNA ...our blog post,Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening. Also, check out our ...plasmid that was used by Chen and colleagues for genome editing inXenopus tropicalis. (Guo et al., Development...Development. 2014..) A new, higher specificity genome editing system that combines TALENs and CRISPRS. Developed...
  11. Supporting Rare Disease Research with the 2020 BeHEARD Awards

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    Blog Post
    ...The Rare Genomics Institute BeHEARD (Helping Empower and Accelerate Research Discoveries) Challenge provides... year is our fourth year working with the Rare Genomics Institute by providing plasmid-based tools for...Additional Resources Find out more about the Rare Genomics Institute and this year's BeHeard Award! ...
  12. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...component that physically integrates into the host genome, the virus is replication incompetent, meaning ...humans and does not always integrate into the host genome as part of the viral infection process (though ...AAV in that it does not integrate into the host’s genome. However, some adenovirus strains are known to ...
  13. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...especially useful within both the synthetic biology and genome engineering fields. We’ll walk you through how ...website or Engler & Marillonet. Golden Gate and genome engineering In early 2011, the Bogdanove and Voytas...described a new Golden Gate-based technology for genome editing which allowed for the ordered assembly ...
  14. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...the worm genome and make additional mutations in our worms.  We are on the lookout for genome modifications...molecular biology are based on the standard worm genome. While C. elegans are often thought of as simple...
  15. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...development of phosphoserine incorporation Extensive genomic sequencing of prokaryotic lineages revealed a curious...effects on cell growth. The C321.ΔA strain was genomically recoded to eliminate all 321 amber stop codons...Pirman, Natasha L., et al. "A flexible codon in genomically recoded Escherichia coli permits programmable...PMCID: PMC4566969. 7. Lajoie, Marc J., et al. "Genomically recoded organisms expand biological functions...
  16. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ...samples in our queue such as viral samples for Viral Genome Sequencing (VGS). QC is performed throughout the...yield sufficient data to capture the entire viral genome. The full details of that workflow and its analysis...Platt RJ, Brigham MD, Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9 knockout and transcriptional ...
  17. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...sensors, our approach uses portions of the pathogen’s genome as unique barcodes to identify samples containing... Of the 48 sensors, targeting 24 regions of the genome, we have deposited the best performing sensors ...encoding corresponding portions of the Zika virus genome with Addgene to facilitate rapid sharing of these... Sample 6 Cofounder Michael Koeris Learn How Minigenomes Are Being Used to Study Dangerous Viruses Resources...
  18. Editor's Choice, September 2016

    Type
    Blog Post
    ...useful to your colleagues. Technique: Probe Phage Genomes for Host Binding Proteins & Running for Rare Disease...this month. The first, "Technique: Probe Phage Genomes for Host Binding Proteins," was contributed by ...
Showing: 221 - 240 of 534 results