Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 535 results for: genome

Showing: 161 - 180 of 535 results

  1. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...integrates into the genome, CRISPR may increase this rate. Specifically, portions of the AAV genome can integrate...is the process of making a hybrid AAV, with the genome from one serotype and the capsid of another. Researchers...delivered by AAV does not integrate into the host genome. AAV instead relies mainly on episomal expression...pieces of DNA that reside outside a host cell’s genome. In this state, AAV cargo can be stably expressed...Overcoming the AAV Size Limitation for CRISPR Delivery Genome-wide Screening Using CRISPR Performing In Vivo ...

  2. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...since those will quantify both genome-containing (full) and non-genome-containing (empty) capsids, thus...concentration of viral particles containing a viral genome. While ddPCR is superior in precision and variability... we recommend using primers targeting the viral genome or inverted terminal repeats (ITR). You should ...particles via a novel capsid ELISA: Packaging of genomes can limit production of recombinant AAV-2. Gene...self-complementary adeno-associated viral vector genome titers by droplet digital PCR. Human Gene Therapy...

  3. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...Results with ICE on Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing...delivery of CRISPR components into target cells, genomic DNA from both edited and unedited (control) populations...editing. ICE software identifies the percentage of genomes that have been successfully modified with insertions...indel sizes in the entire edited population of genomes. Hovering over each bar of the indel plot shows...insertion or deletion along with the percentage of genomes that contain the indel. The discordance plot (right... analysis tool is part of Synthego’s full stack Genome Engineering Solutions, which also includes our ...

  4. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...detect chromatin interactions at one location in the genome at a time (Liu et al., 2017). Recently the Xu lab..., a biotin ligase a gRNA(s) targeting a single genomic location of interest  When these three components...that a new cell line has to be created for each genomic loci of interest. This is cumbersome and also makes...3C-based techniques: the powerful tools for spatial genome organization deciphering. Molecular Cytogenetics...locus-specific 3D chromatin by biotinylated dCas9. Genome Biology, 21. https://doi.org/10.1186/s13059-020...Addgene.org Find Other Cas9 Variants for Purifying Genomic Regions of Interest Here Brush Up on Your CRISPR...

  5. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...kb T7 phage genome. They first compared 10-piece linear (the native state of the T7 genome) and circular...tools, they successfully assembled a 40 kb T7 phage genome from 52 (!!) fragments in one very Golden (Gate... ones. The 52-piece circular assembly of the T7 genome had lower efficiency than the 10-piece circular...possible to select for correctly assembled phage genomes, it required a very stringent selection process...Pokhrel, N., & Lohman, G. J. S. (2022). Rapid 40 kb Genome Construction from 52 Parts through Data-optimized...

  6. Evolution of Lab Techniques

    Type
    Blog Post
    ... at specific sites (5). Imagine that you have a genome with unknown sequence. In the era before sequencing...mapping was commonly used to map DNA and examine genomes (6). The knowledge gained through these experiments...revolution that still prevails. Sequencing whole genomes or profiling transcriptomes (RNAseq) require sequencing...soon become obsolete. For instance, the popular genome-engineering tool, CRISPR, may one day make techniques...sequencing technologies could one day sequence an entire genome for under $1,000 (10). The future of sequencing...of sequencers: The history of sequencing DNA." Genomics (2015). PubMed PMID: 26554401. PubMed Central ... fourth-generation DNA sequencing technology." Genomics, proteomics & bioinformatics 13.1 (2015): 4-16...

  7. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...  CRISPR knockout screens: Full genome vs. custom screens Full genome CRISPR knockout screens have been... are used for each gene. At the moment, 13 full genome SpCas9 knockout libraries designed by different... a major limitation. An alternative to the full genome CRISPR screen approach is to use smaller libraries...generated a list of candidate genes from a full genome screen, a smaller, secondary, CRISPR screen could...Green Listed, as of now, contains most of the full genome knockout human and mouse libraries available through...Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide libraries for CRISPR screening." Nature methods.... 7. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library...

  8. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...for introducing one modification into the mouse genome, they are not commonly used to introduce multiple...generate mice that have two modified genes in their genome, and assuming Mendalian patterns of inheritance...integration of a novel gene randomly into their genome, some transgenic strains are difficult to breed...posts Read blog posts about using Cre-lox Read our genome engineering blog posts Resources on Addgene.org...Addgene.org Find more about Cre-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...

  9. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...copies that have integrated into the target cell’s genome. Physical titer The two most common assays for ... has had one viral particle integrated into its genome and a cell that has had multiple integration events...copies that have integrated into a target cell’s genome. This approach can discriminate multiple integration...transduced with serial dilutions of a viral prep, genomic DNA isolated, and primers targetting viral components...Lentivirus FAQ Learn How Lentivirus is Used in CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used...

  10. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ... editors create specific point mutations in the genome, but they’re inefficient compared to CRISPR/Cas9... and to target specific loci of interest in the genome. The Wang and Brafman lab showed that GFP positive... TREE have a significantly higher frequency of genomic base pair edits of interest compared to cells segregated...adenine base editors, that make A-to-G edits in the genome (Gaudelli et al., 2017). GO is thus a flexible ...2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . ... for the Quantification of CRISPR/Cas9-mediated Genome Editing. Molecular Therapy - Nucleic Acids 5:e334...

  11. Viral Vectors 101: Transductions

    Type
    Blog Post
    ...twist is that the typical genetic cargo — the viral genome — is swapped, at least partially, for a nucleic...other cells, but since the majority of the viral genome has been swapped for alternative genetic cargo,...nucleic acid cargo to be integrated into the host genome? Expressed episomally long-term? Expressed short-term...For example, some viruses integrate into the host genome (lentivirus and retrovirus) while others are non-integrating...

  12. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...coupled to high-throughput sequencing to identify genome-wide binding patterns of the target protein. Both...Transcript ID(s) of your favourite gene(s) from Ensembl genome browser: https://www.ensembl.org/index.html Design...polymerase II for transcription) are more effective for genome-editing events. In the light of these results, ...now, the tool is available for human and mouse genomes only and deals with C-terminus epitope [email protected]. Keep pipetting and happy genome editing! Pooran Dewari is a postdoc in Steve Pollard...research interests include optimizing CRISPR/Cas9 genome-editing methods and applying them to study brain...JD. 2017. Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA...

  13. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...SpCas9’s 3’ NGG PAM occurs frequently in GC-rich genomes, but a PAM is not always available near the locus...there is an NGG PAM every 42 bases in the human genome. SpCas9 targeting should be easy, right? Yes and...TTTV PAM, improve targeting in AT-rich loci or genomes. Cas9s from other species, like S. aureus and N...more complex than NGG, resulting in lower overall genome frequency. Despite the progress we’ve made, there...al. performed GUIDE-seq analysis to characterize genome-wide off-targets, since increasing PAM flexibility...

  14. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...evolving. Variants of Cas proteins can be used for genome editing, activating gene expression, repressing...CRISPR system, N. meningitidis can also be used for genome editing. In this study, the labs found that anti-CRISPR... Conclusion CRISPR is a great way to make those genome edits. But, just as important is a way to flip ...Peng X (2018) Anti-CRISPR-Based and CRISPR-Based Genome Editing of Sulfolobus islandicus Rod-Shaped Virus... genome editing...

  15. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ... crosses. Fly genomes Compared to humans and mice, flies have a relatively simple genome. They have four...gene? Can you find an ortholog in the Drosophila genome? Continue reading and I’ll show you how Drosophila...are diploid except for their germ cells (sets of genomic material are contributed by mother and father)....live animals yet, Harvard’s DRSC/TRiP- Functional Genomics Resources website has in vitro reagents and tools...

  16. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...2016 was an exciting year for genome engineering research. A variety of new tools came out including ...SpCas9 is still going strong as the basis for many genome editing experiments. So, without further ado, the...describing them here. References 1.Ran, F. Ann, et al. "Genome engineering using the CRISPR-Cas9 system." Nature...

  17. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...transcriptional activators to induce gene transcription at genomic sites targeted by specific sgRNAs. Its RNA-guided...nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries. However, using... the p65-HSF1 transcriptional activator to each genome-bound dCas9 (see Figure 1a for the transgenic construct...Because CRISPRa can be targeted anywhere in the genome, it is relatively straightforward to use CRISPRa...occur. CD8+ cells were then isolated and their genomic DNA sequenced. Guide RNAs targeting several known...demonstrated by the Qi Lab, which developed a subgenome-scale CRISPRa screening approach in embryonic ...

  18. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ..., amplified the CRISPR locus from the bacterial genome, and inserted it into E. coli to express and then...worth of DNA with CRISPR-driven precision into a genome, can be considered the fourth generation of CRISPR...fusions, using Prime to lay a “beacon” into the genome that a fused integrase would then integrate into...10-20% integration of inserts up to 36 kb into a genome. This integration was observed in non-dividing ...S. Gootenberg, Omar O. Abudayyeh. Drag-and-drop genome insertion without DNA cleavage with CRISPR-directed...

  19. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ... end up with a titer of 4 x 1012 genome copies/mL or 8 x 1012 genome copies/mL and both results would ...dynamic range of the ddPCR is between 1 and 100,000 genome copies (GC) per reaction. Since AAV titers tend...can be used to examine the integrity of the viral genome (Furuta-Hanawa et al., 2019). Tips and tricks for...self-complementary adeno-associated viral vector genome titers by droplet digital PCR." Human gene therapy...
Showing: 161 - 180 of 535 results