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We narrowed to 535 results for: genome

Showing: 181 - 200 of 535 results

  1. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...packaging to infiltrate and integrate into the host genome. First, it needs to be virally enveloped which ... and permanently insert the cargo into the host genome. Lentiviral-specific components Lentiviruses have...While AAV often does not integrate into the host genome (more on this below), it’s still used to deliver...for viral replication and also aids in infrequent genome integration at the AAVS1 locus in humans. Cap is...

  2. Plasmids 101: Screens vs. Selections

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    Blog Post
    ...small molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with gRNA libraries...and then outcomes are measured. While the entire genome can be knocked out with CRISPR, the entirety of...knocking out the genes of interest (or the whole genome), they knock down gene expression temporarily. ...resources on the Addgene Blog CRISPR libraries Genome-wide screens with CRISPR Lentiviral CRISPR libraries...

  3. Hot Plasmids - February 2022

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    Blog Post
    ...Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing Anti-FLAG Frankenbodies...Cas12a-based screening method was used to create a genome-wide library capable of performing similarly to...Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing by: Rachel Leeson The large...applications.  Figure 4: Overview of CasMINI for genome engineering applications. Image from Xiaoshu, et...

  4. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...understand the function of the non-coding portion of the genome. Mendenhall and Richard Myers of HudsonAlpha (where...define all of the functional elements in the human genome. The pair has focused considerable attention ... gene regulatory proteins and the places in the genome where they can be found more quickly and efficiently...transcription factors and figure out where they bind to the genome. We’ve been doing that for years using this assay...

  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...from a random genomic location than to express the tagged protein from a defined genomic location or from...FRB-NES) resulting in a rapamycin-inducible Cas9 for genome editing. Without rapamycin treatment, the Cas9(...greater temporal control over CRISPR/Cas9 mediated genome modification and gene expression. The plasmids ...has been targeted to a very specific site on the genome via the CRISPR system. The ease and flexibility...reporter gene, or (4) multiple sgRNAs with Cas9 for genome engineering. The versatility and ease of generating... Multicolor CRISPR labeling chromosomal loci in genome Thoru Pederson’s lab at the University of Massachusetts...tool enabling the imaging of multiple endogenous genomic loci simultaneously in live cells. This system ...

  7. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...method for in situ germline genome engineering using CRISPR nucleases. Genome Biol. 19 (2018) (1):25. DOI...CRISPR genome editing has made it easier to create knockout alleles in a variety of species, including...developed i-GONAD as a simple method for delivery of genome editing components to the germline. In contrast... and conditional alleles by CRISPR/Cas-mediated genome engineering." Cell 154 (2013):1370-9. DOI: 10.1016...long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186/s13059-017...

  8. RaPID Detection of RNA-protein Interactions

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    Blog Post
    ...plasmids for labeling genomic regions Find CRISPR plasmids for purifying specific genomic regions  ...the literature and analyzing data from The Cancer Genome Atlas, Ramanathan et al. found evidence that suggests...experimental identification of RNA-protein interactions. Genome Biology. PubMed PMID: 24467948 PubMed Central PMCID...

  10. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...uses fluorescently labeled Cas9 to label multiple genomic loci in live cells. While other systems can be ...is a great step forward in live cell imaging of genomic loci. Ma et al. anticipate that the technique will...this publication at Addgene. Dynamic imaging of genomic loci in living human cells by an optimized CRISPR...

  11. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...modifying capsid and genomic structures. For example, rAAV pseudotypes mix the genome of one AAV serotype... AAV vectors have both distinctive capsids and genomic structures. The viral capsid is the key that lets...separated by a slash: the first number refers to the genome, and the second number to the capsid. Most vectors... provided by Addgene are pseudotypes using the genomic structure of AAV2, so they are formally called ...best studied and most commonly used serotype. Its genome is the most common choice for AAV pseudotyping....

  12. Quick Guide to All Things Lentivirus

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    Blog Post
    ...broader range of cells and their integration into the genome is more efficient. The first retroviral vectors...three plasmids encoding 1) the lentiviral vector genome which was composed of the wild-type 5’ and 3’ LTRs...desired gene (transfer vector plasmid), 2) the HIV-1 genome containing all viral genes with the exception of...LTR so you don’t need Tat to transcribe the viral genome in producing cells. Currently, the third-generation...still infect the researcher and insert into her/his genome - this is a biosafety risk Infects dividing and...proteins generated Stable integration into host genome and stable expression of the transgene   Transfer...

  13. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...stable fly cell lines; attL and attR sequences for genome integration by phiC31) Drosophilia Plasmid...fly cell lines; Hygromycin resistance gene; for genome integration by spontaneous insertion)   Drosophilia...sequences; white + selectable marker; attB site for genome integration by phiC31   The Fragmid website...assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/j.xgen.2024.100519...

  14. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

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    Blog Post
    ...loci – in neighboring chromatin or throughout the genome – could be affected, leading to unpredicted gene...targeting, including modifying other sites in the genome and directly interacting with transcription machinery...DNMT1 from that site. So, when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites...Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints...Resources on the Addgene blog CRISPR101: Editing the Epigenome  Mapping the 4D Nucleome with CRISPR/Cas9 Resources...

  15. CRISPR in the Clinic

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    Blog Post
    ...Cas9 technology has been a breakthrough for the genome-editing field and the greater scientific community...several strains of E. coli and lethally chop their genomes into many, many pieces. Locus Biosciences is currently...employs Cas9 to cut at two sites within the viral genome to excise a large portion of it. This ongoing trial... Zarghamian P, Klermund J, Cathomen T. Clinical genome editing to treat sickle cell disease-A brief update...

  16. CRISPR 101: Mammalian Expression Systems and Delivery Methods

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    Blog Post
    .... CRISPR technology has been widely adopted for genome editing purposes because it's cheaper, faster, ...be used in vivo. A common choice for conducting genome-wide screens using CRISPR. AAV transduction Only...evaluate lentiviral component delivery as well as genome repair by non-homologous end joining (NHEJ) or ...component delivery and sgRNA efficacy by cloning in genome target sequence into EGFP reporter. Target DNA ...

  17. Typing CRISPR Systems

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    Blog Post
    ...2022b). Genome editing technology and applications with the type I CRISPR system. Gene and Genome Editing...CRISPR, by any other name would…probably still edit genomes. Understanding the different types of CRISPR systems...might imagine, this is handy for creating large genomic deletions. Type I systems are also employed as ...Marraffini, L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/CAS systems. Science, ...inactivation mechanisms in type I-C CRISPR-Cas3 for genome-editing applications. Molecular Cell, 84(3), 463... expanding biotechnology toolbox for and beyond genome editing. Cell & Bioscience, 8(1). https://doi.org...

  18. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

    Type
    Blog Post
    ...profiling samples, and years later, several whole genome sequencing projects were launched in the Quake ... Schmidt not only produced partially assembled genomes, but also led to the discovery of IgnaviCas9, a...CRISPR array in the resulting Ignavibacteriae phylum genome. IgnaviCas9 is a hyperthermostable Cas9 CRISPR ...Spring in Lower Geyser Basin. They used a mini-metagenomics method using microfluidic parallelization to...

  19. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    Blog Post
    ...and the transfer plasmid which contains the viral genome. In today’s blog post, we’ll focus on the AAV ...transgene. The WPRE is located at the end of the AAV genome, just upstream of the final ITR.  polyA  The polyadenylation...longevity. The polyA is located at the end of the AAV genome, just upstream of the final ITR and downstream ...

  20. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...model generation CRISPR has made the generation of genome edits accessible, fast, and easy. In cases where...Line Using CRISPR-Cas9 to Edit Disease Out of the genome Tips for a first time CRISPR User (by a first ...and Resources CRISPR History and Development for Genome Engineering    ...
Showing: 181 - 200 of 535 results