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We narrowed to 798 results for: genome

Showing: 221 - 240 of 798 results

  1. 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 ...

  3. Gibson Assembly Protocol

    Type
    Protocol
    .... Chemical synthesis of the mouse mitochondrial genome. Nature Methods , 7(11), 901–903. https://doi.org...

  4. An Introduction to Adenovirus

    Type
    Blog Post
    ...viruses with a recombinant genome, and adenoviral vectors all have a genomic capacity that is much higher...non-enveloped double-stranded DNA viruses, with a linear genome between 26–45 kb in length. Human adenovirus type..., gene expression, and protein translation. Its genome consists of “early” (E) and “late” genes, and the...late gene transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early...cell line used (which have fragments of the AdV genome in them, including the E1 region). The space in...that is made available by this deletion from the genomic plasmid can be used to house a recombinant sequence...adaptive immune responses that inactivate viral genomes or otherwise clear the virus from the cell population...

  5. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...introducing double stranded breaks (DSBs) into genomic DNA in many cell types and often results in biallelic...sequence changes to be incorporated into the edited genome. In mammalian cells such as stem cells, HDR is ... HDR rates, we have shown recently that desired genome-editing events by HDR can be generated more efficiently...accuracy with CRISPR/Cas-blocking mutations After genome editing with CRISPR/Cas9 in human cells was described....     References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." Science ...PMC3795411. 2. Mali, Prashant, et al. "RNA-guided human genome engineering via Cas9." Science 339.6121 (2013):..., Dylan, et al. "Precise and efficient scarless genome editing in stem cells using CORRECT." Nature protocols...

  6. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... Get the Basics of Mouse Genome Engineering with CRISPR Validate Your Genome Edit Learn about Non Homologous...mice? We'll have a post that goes into the mouse genome editing process in a bit more detail in the coming... where you are looking, you can either scan the genome yourself for potential gRNA sites, or you can utilise...ubiquitous CAG promoter. The approximately 500 bp genomic fragment containing the sgRNA target sequence can...researchers first started using CRISPR/Cas9 for genome editing in mice, they would often microinject gRNAs...Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1 (2014): 132-141. PubMed PMID: 24253446...mutations in multiple genes by CRISPR/Cas-mediated genome engineering." Cell 153.4 (2013): 910-918. PubMed...

  7. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    ...of the exciting developments surrounding CRISPR genome engineering in the past year, we're excited to ...library screening, read our recent blog post on Genome-wide Screening Using CRISPR/Cas9. If you need help...sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening in human cells...Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide libraries for CRISPR screening." Nature methods...

  8. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others use...different plasmids. Harvard’s DRSC/TRiP Functional Genomics Resource Center has various plasmids for overexpression...color depends on how transcriptionally active the genomic locus of integration is. The white gene is most... Learn about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific... genome engineering...

  9. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...off-target activity. Deleting a large region of the genome by removing the sequence between two target sites...multiplexed gRNAs can target multiple locations in the genome to modify, whether it be editing CRISPRi, CRISPRa...gRNA expression, and thus the highest frequency of genome editing events. Yamamoto Lab Multiplex CRISPR/Cas9...hypopigmentation, thus marking cells that have undergone genomic modification. In this system, Cas9 must be supplied... recombineering-based MAGE (Multiplex Automated Genome Engineering) technology. pMA7CR_2.0 expresses lambda...Kabadi, Ami M., et al. “Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector.” Nucleic...Addgene. 3. Sakuma, Tetsushi, et al. “Multiplex genome engineering in human cells using all-in-one CRISPR...

  10. The 12 Days of CRISPR: 2021

    Type
    Blog Post
    ...Precise genomic deletions).  On the seventh day of CRISPR, Addgene gave to me: programmed genome editing... the PEDAR strategy to delete and replace long genomic sequences. On the ninth day of CRISPR, Addgene ...

  11. 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...

  12. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...NgAgo and showed that it can cleave plasmid and genomic DNA in a variety of mammalian cell lines, with ...should have much more flexibility in choosing a genomic target. NgAgo also appears to outperform Cas9 in...hypothesized that base removal helps prevent the invading genome from “recovering” its original sequence. If you...Feng Jiang, Yongqiang Wu & Chunyu Han. “DNA-guided genome editing using the Natronobacterium gregoryi Argonaute... Check Out High Fidelty Cas9 Mutants Browse All Genome Engineering Posts Additional Resources on Addgene.org...

  13. The 12 Days of CRISPR

    Type
    Blog Post
    ...Precise Genome Editing (CRISPEY!)🎵 On the 7th day of CRISPR, Addgene gave to me: Drag-and-drop genome insertion...me: dCas9-based gene editing for cleavage-free genomic knock-in of long sequences🎵On the 2nd day of CRISPR...

  14. 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...

  15. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...Prime editing is a versatile genome editing technology that allows precise modifications of DNA (replacements...substantially affects prime editing outcomes across many genomic locations. In our study, we used the TRIP (Thousands...efficiency to chromatin data from over a thousand genomic locations. This allowed us to train ePRIDICT (Figure...(Figure 3), a model that predicts how well a genomic site can be edited based on its chromatin environment...Raguram, A., & Liu, D. R. (2019). Search-and-replace genome editing without double-strand breaks or donor DNA... Prime editing for precise and highly versatile genome manipulation. Nature Reviews Genetics, 24(3), Article...Addgene blog How to Design Your gRNA for CRISPR Genome Editing PRIME Editing: Adding Precision and Flexibility...

  16. Viral Production at Addgene

    Type
    Blog Post
    ...sequencing of the Viral Genome Next-generation sequencing is performed on viral genomes isolated from the final...confirm the identity and integrity of the viral genome and the absence of unexpected DNA contaminants....pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] ...Electron Microscopy The ratio of empty to full (i.e., genome-containing) AAV particles within representative...lentiviral vector, incubated for 72 hours, and genomic DNA is extracted. Primers and probes targeting ...self-complementary adeno-associated viral vector genome titers by droplet digital PCR. Human Gene Therapy...

  17. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ... ability to integrate permanently into the host genome and thereby enable long-term stable gene expression...MuLV, or MLV) or Murine Stem Cell Virus (MSCV) genomes whereas lentiviral vectors are derived from the...the human immunodeficiency virus (HIV) genome. For research purposes, retroviral vectors have been developed...allow functional screening of thousands of genes in genome-wide experiments. Gamma-retroviral vs. lentiviral...support functional investigations of the mammalian genome that can reveal how genetic alterations lead to... CRISPR and the most recent developments in the genome editing field. Lentiviral Plasmid Kit Multiple...constantly improving our CRISPR reagents. Please check www.genome-engineering.org for the most up-to-date information...

  18. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021)....to mammalian cells (Allen, et al., Frontiers in Genome Editing).  Some of the RNA stabilizing modifications... gRNA introduction (Allen, et al., Frontiers in Genome Editing). This is particularly important in experiments...Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol ...for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation. ACS 6, 5, 695-703 (2020). https://pubs.acs.org...

  19. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...in the genome near either the beginning or end of the gene of interest —in our case in the genome of an...inserting large tags into a precise location in the genome is still inefficient, particularly in human cell...plasmids work and to discover more resources for your genomic tagging experiments. Browse the Allen Institute...an hiPS cell. While the majority of the time the genome will attempt to repair itself without outside influence...make your own, will make it easier for you to use genomic tagging in your experiments. Many thanks to our...

  20. 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...
Showing: 221 - 240 of 798 results