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We narrowed to 535 results for: genome

Showing: 121 - 140 of 535 results

  1. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...homologous recombination. Modifying bacterial genomes using exogenous enzymes: Lambda red and CRISPR ...generate off-target mutations. Modifying bacterial genomes using counter-selectable markers and homologous...recombination enzymes can also accomplish the desired genomic modification. This technique is frequently called...undergone single crossover and integrated into the genome. Subsequent selection on sucrose will identify ...technique for the scarless modification of bacterial genomes. It can accomplish large deletions, insertions,...Precision-engineering the Pseudomonas aeruginosa genome with two-step allelic exchange." Nature protocols...Wenyan, et al. "RNA-guided editing of bacterial genomes using CRISPR-Cas systems." Nature biotechnology31.3...

  2. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...exchange. By utilizing recombinases to manipulate genomes, scientists can control expression of their genes...pubmed.ncbi.nlm.nih.gov/24390935/ Hypercompact genome editor CRISPR-Casɸ By Melina Fan The Doudna Lab... their discovery of CRISPR-Casɸ, a hypercompact genome editor from the Biggiephage clade of huge phages...Doudna Lab. Scientists looking for more compact genome editors due to delivery constraints and scientists...scientists seeking a wider range of targetable genomic sequences could benefit from CRISPR-Casɸ. Pausch et...CRISPR screens by using this lentiviral CRISPR human genome-wide library deposited by Leopold Parts. This library...any reduction in performance compared to larger genome-wide libraries. Find the library here New from...

  3. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ... the construct has randomly integrated into the genome, a negative selection marker like the HSV thymidine...incorporate the negative selection marker into the genome. This final product of non-homologous recombination...knock-in, any gene, tag, or mutated exon into the genome. For this purpose, the sequence to be inserted ...both knockout a given gene and insert GFP into the genome, we’d create a plasmid similar to the one shown...ranging from undetectable to 0.1%. In contrast, new genome editing techniques such as CRISPR are easier to...efficient at inactivating genes. CRISPR can target a genomic sequence and create a break that can be repaired...cells and mouse knockout lines should increase. New genome engineering tools also offer hope for the creation...

  4. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...degradation, and proliferation. Genome manipulation K. phaffii’s genome has been sequenced for some time...yeast is a powerful eukaryotic model organism – its genome is easy to manipulate, it’s affordable, and it ...The first eukaryote (S. cerevisiae) to have its genome fully sequenced There are several human disease...time, clearing the greatest hurdle for genome engineering. A unique evolutionary feature of K. phaffii,...fmicb.2020.607028  Riley, R., et al., Comparative genomics of biotechnologically important yeasts. Proc. ...

  5. 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...

  6. 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...

  7. 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...

  8. 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...

  10. Gibson Assembly Protocol

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

  11. 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...

  12. 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...

  13. 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 ...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  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. 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...

  20. 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...
Showing: 121 - 140 of 535 results