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We narrowed to 411 results for: genome

Showing: 121 - 140 of 411 results

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

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ...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...

  2. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...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...

  3. The 12 Days of CRISPR: 2021

    Type
    Blog Post
    Published
    Dec. 28, 2021, 2:15 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...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...

  5. The 12 Days of CRISPR

    Type
    Blog Post
    Published
    Jan. 3, 2023, 2:15 p.m.
    ...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...

  6. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...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...

  7. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...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...

  8. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...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...

  9. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ... 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...

  10. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ...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...

  11. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...mechanisms. The RNA guide is typically 20 nts and the genomic locus must often be proximal to a PAM site for ...gRNA with low sequence similarity elsewhere in the genome. There are many predictive online tools which can...gRNA with low sequence similarity elsewhere in the genome to reduce off-targets, it is time to consider your...Even if there are similar sequences present in the genome, certain Cas enzymes are less compatible with the...several options to quantify these events. Whole genome sequencing The only way to quantify off-target ...off-target effects entirely is to perform whole genome sequencing. While this may be unnecessary for many applications...gRNA which doesn’t perfectly match any site in the genome. While this may effectively serve as a transfection...

  12. What's New in CRISPR - December 2019

    Type
    Blog Post
    Published
    Dec. 17, 2019, 2:37 p.m.
    ...for the Application of Neuronal Genome Editing) mediates targeted genomic integration of fluorescent tags... editors, which decrease the rate of off-target genome edits compared to rat APOBEC1. Now, they’ve engineered... developed a CRISPR-based toolkit for targeted genomic integration of fluorescent tags in neurons. Their...

  13. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only transduction? Niels Geijsen...see if iTOP would be compatible with CRISPR/Cas9 genome editing. Although CRISPR/Cas9 is all the rage, ...mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient, which... these reasons, iTOP, especially in tandem with genome editing, may make primary cell work much more efficient... genome editing...

  14. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...and accessible genome editing tool.   References Mangeot, Philippe E., et al. "Genome editing in primary...CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability...) virus-like particles (VLPs). VLPs lack viral genomes which means they are non-infectious and non-replicative...Nanoblades? Plasmids are available from Addgene. Genome editing primary cells with Nanoblades Primary cells...

  15. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    Published
    Sept. 9, 2016, 2:43 p.m.
    ...scale. Or you don’t have enough cells to execute genome-wide perturbations. So, while some might argue ...mouse and human kinomes. While any subset of the genome is limited by definition, the kinome is a well-...as opposed to the 70,000 or more gRNAs found in genome-wide libraries, and thus can be screened in pooled...Resources on the Addgene Blog Learn to Conduct Genome-wide Screens with CRISPR/Cas9 Get John Doench's...

  16. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...cataloged in the ClinVar database. Previously developed genome editing methods using nucleases and base editors... published in Nature, is a “search-and-replace” genome editing technique that mediates targeted insertions...also encodes the desired edit and homology to the genomic DNA locus. After nicking by Cas9, the homologous...location The prime editor extends the reach of CRISPR genome editing, as it can edit near to or far from PAM... Fyodor Urnov refers to as “PAM deserts” in the genome. More versatile and precise than base editing (...achieve the best efficiency. The type of edit, genomic context, delivery strategy, cell type, and goal...(though up to 45-fold) higher across different genomic sites compared to PE1 (Anzalone et al., 2019). ...

  17. Introducing Addgene's Microbiology Pages

    Type
    Blog Post
    Published
    June 13, 2016, 7:08 p.m.
    ...Check out our Genome Engineering Overview and our CRISPR guide to find out how to use genome editing techniques...explaining how CRISPR is being used for bacterial genome engineering. To make it easier for you to find...

  18. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...our genome is made of protein-coding genes, and you might be wondering what the rest of our genome could...splice sites. (C) Removal of an exon or a large genomic fragment. (D) Knock-in: insertion of a synthetic...with specific chromatin structures or other local genomic elements (Hazan and Bester 2021). Other Cas variants...doi:10.1038/s41587-020-0428-0 Rinn JL, Chang HY. Genome Regulation by Long Noncoding RNAs. Annual Review...annurev-biochem-051410-092902 Liu Y, Cao Z, Wang Y, et al. Genome-wide screening for functional long noncoding RNAs...

  19. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...transferred, Mobile-CRISPRi stably integrates into the genome and is propagated during normal DNA replication...element is first introduced into the B. subtilis genome. Mobile-CRISPRi is then transferred from B. subtilis...mating. Following excision from the B. subtilis genome, the Mobile-CRISPRi ICE element forms a conjugative... downstream of the glmS gene in the recipient's genome. Antibiotic plates lacking DAP select against donor...colonies. Higher transfer rates are better suited for genome-wide CRISPRi library screening experiments where... Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi...

  20. 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 . ...
Showing: 121 - 140 of 411 results