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We narrowed to 411 results for: genome

Showing: 141 - 160 of 411 results

  1. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...that targets the complex of TadA and dCas to the genomic DNA of interest  Figure 1. When ABEs are... pyogenes Cas9 requires the target site in the genomic DNA to contain a specific protospacer adjacent ...ABE8 could target many additional locations in the genome. Compared to ABE7.10, NG-ABE8.xm/d showed a 1.6x... 8 selected ABE8s and ABE7.10. They quantified genomic editing by measuring the reduction in protein expression...note, ABE8s reported in this work did not cause genome-wide, guide-independent off-target deamination,...2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . ...

  2. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...complementary to the target site, or protospacer, in the genome. Native Cas9 has two catalytic domains, each of...of gene expression is observed in the absence of genome editing (Fig. 2 and 3) (5, 3). Figure 2:.... 2. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library...Kiani, Samira, et al. "Cas9 gRNA engineering for genome editing, activation and repression." Nature methods...PMC4666719. 4. Mali, Prashant, et al. "RNA-guided human genome engineering via Cas9." Science 339.6121 (2013):...Randall J., et al. "CRISPR-Cas9 knockin mice for genome editing and cancer modeling." Cell 159.2 (2014)...

  3. CRISPR Cheat Sheet

    Type
    Blog Post
    Published
    Jan. 27, 2021, 2 p.m.
    ...bacterial immune system. Genome Editing: Modifying the DNA sequence of an organism’s genome usually for a research... genome editing...

  4. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    Published
    Sept. 15, 2014, 6:05 p.m.
    ... will be bestowed upon discovers of CRISPR/Cas9 genome engineering, Dr. Jennifer Doudna and Dr. Emmanuelle...and its potential applications in rewriting the genome. Their award acceptance addresses were inspiring...discussed how J&J is already modifying cell and animal genomes to test hypotheses of disease. They are finding...more about safety and more about the how to use genome edited stem cells to treat disease. As for me, ...

  5. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ...set of problems. Rapidly dividing cells initiate genome replication more than once per cell division and...constitutive promoters drove maximal gene expression near genomic origins of replication, the stabilized promoters...equivalent whether the genes were expressed from the genome or a plasmid. Thus, a stabilized promoter can allow...allow a system to be moved from a plasmid to the genome without greatly impacting the level of gene expression...constitutive promoter and then moved them into the genome, deoxychromoviridans production titers decreased...

  6. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ...into the genome and stable cell lines, where foreign DNA is incorporated into the cell’s genome and is ...genetic material is incorporated into the host’s genome at a particular integration site. The integrated..., or it can integrate into the host bacterium’s genome through homologous or site directed recombination...sequences of interest into a bacterial cell or a host’s genome. To do this scientists commonly use phagemids, ...

  7. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ...genetic payload. This cargo is typically their own genome, but this step can be engineered to be anything... then be either permanently integrated into the genome of the target cell or transiently expressed. It...higher than traditional transfection and random genomic integration of a plasmid. Viral transduction is...These libraries include sgRNAs targeting the entire genome, overexpression of all human transcription factors...Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells Additional Resources ...

  8. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ...good. RNA:RNA dsRNA and viral infection Viral genomes come in all sorts of shapes and sizes. Many are...mitochondria. These organelles have their own circular genomes which can produce dsRNA as a result of bidirectional...regions, such as telomeres and centromeres. These genomic regions are prone to persistent R-loop formation...damage, and these structures can in turn lead to genome instability. Thus, it’s important to strike a balance...Human proteins that interact with RNA/DNA hybrids. Genome Res., 28(9): 1405-1414 (2018). 10.1101/gr.237362.118...

  9. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...the short time since its development, CRISPR/Cas9 genome editing has been used to study the effect of gene...expression of gRNAs from these constructs could lower genome editing efficiency. One previous strategy for... rice protoplasts. Each pair of gRNAs targeted genomic sites separated by 350-750 bp, allowing them to...CRISPR/Cas9 shows great potential for multiplexed genome editing. Interested in using PTG plasmids in your...CRISPR RNA-guided FokI nucleases for highly specific genome editing. Tsai SQ, Wyvekens N, Khayter C, Foden ...

  11. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...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 ...

  12. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...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...

  13. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Aug. 1, 2023, 1:15 p.m.
    ...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...

  16. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ... 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...

  17. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...  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...

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

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...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 ...

  19. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ... 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...
Showing: 141 - 160 of 411 results