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We narrowed to 704 results for: des.1

Showing: 161 - 180 of 704 results

  1. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...disadvantages: Table 1 (Joglekar et al., 2017)  Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al...polyA tail.   Figure 1: Lentiviral production uses three plasmids: (1) The transfer plasmid, (2)...can be used for specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020). Because...pseudotyping allows these viruses to infect neurons. Besides altering the host tropism of your virus, there ...neurons. However, VSV-G’s broad tropism can be undesired for in vivo studies since there’s the potential...VSV-G is also toxic at high concentrations as described above and is inactivated by the complement system...

  2. AAV Packaged on Request is Here!

    Type
    Blog Post
    ...it, select your desired serotype and volume, and submit your request.    Figure 1: The AAV Packaged..., AAV8, or AAV Retrograde (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL. Please note that all orders will... our on-demand viral service, we expect titers of 1 × 1013 GC/mL for AAV1, AAV5, AAV8, and AAVrg and 7...simply request your plasmid of interest in your desired serotype, place your order when ready, and we’ll...

  3. Deep Dive: qPCR

    Type
    Blog Post
    ... Table 1: Features of various qPCR chemistries. Adapted from Wong, 2018   Primers Design of primers...relatively easy to multiplex (Thellwell, 2000). See Table 1 for a comparison of features of several commonly used...show multiple peaks.    (a) (b)  (c)   Fig. 1: Melt curves showsing (a) a single peak, indicating...changes in ΔΔCT are represented by RQs between 0 and 1. Flipping your experimental and control values during...see this fold change as a positive difference (RQ>1), which may be easier to conceptualize. Just remember... in Molecular Biology, vol. 1160, DOI 10.1007/978-1-4939-0733-5_3, © Springer Science+Business Media New... application, Bioinformatics, Volume 27, Issue 7, 1 April 2011, Pages 1019–1020, https://doi.org/10.1093...

  4. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...been designed to perform gene transfer in vertebrates. The system is comprised of 2 components: 1) an ...use is only likely to grow in the coming decades.  References 1. Kowarz E, et al. Optimized Sleeping Beauty...also important for social and regulatory reasons. 1.) The transposon consists of a mirrored set of inverted...T cells to target CD19. Immunol Rev. 2014 Jan;257(1):181-90. doi: 10.1111/imr.12137. PubMed. Find the ...

  5. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ... Figure 1 below) was free to anneal to complementary ssDNA molecules in vitro. Figure 1: Cas9 binding...allowed sequence replacement rates of approximately 1%. This is by no means a large number, but it was attained...who worked on this publication:         References 1. Richardson, Christopher D., et al. "Enhancing homology-directed...DNA post-cut. Using substrate DNA labeled on both sides of the break, we determined that Cas9 held tightly... single or double stranded donor DNA with other design parameters. Example single stranded donor DNA and...approaches. Moreover, the cost of reagents is relatively modest, requiring expressed Cas9 protein, transcribed ... Blog Brush Up on Your HDR Basics Learn How to Design Your gRNA Find the Ideal CRISPR Software for Your...

  6. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...type of assay is a clonal cell line known to contain 1 integrated copy of the qPCR target gene. Developing... successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob Reiser...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009... titration methods."BMC biotechnology 6.1 (2006): 1. PubMed PMID: 16836756. PubMed Central PMCID: PMC1534021...events." Genetic vaccines and therapy 2.1 (2004): 1. PubMed PMID: 15291957. PubMed Central PMCID: PMC514534...virus will not only ensure that your infection is designed in the best possible way, but it may also save...the transgene itself. Many researchers prefer to design primers that target the common features of viral...

  7. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...Correct Path.,” J. stem cell Regen. Biol., vol. 1, no. 1, Jan. PubMed PMID: 26702451. PubMed Central PMCID...genetic manipulation in mammalian cells until 2005 [1]. Like other transposons, piggyBac™ has two components...The predicted frequency of ‘TTAA’ in the genome is 1 in every 256 base-pairs of DNA sequence, making it...has been demonstrated) with no known upper limit [1]. The piggyBac™ technology can be used for numerous...genetic loci of integration [8].   Figure 1: Mechanism of the piggyBac™  transposase/transposon...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-ing models...facilitate the selection of cells containing your desired edit. The selection is marker is then seamlessly...

  8. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...infection, gene targeting vectors with relatively short (1 kb) homology arms underwent homologous recombination...sequences of interest to cells with efficiencies of 1-40% after G418 selection (Kohli et al 2004, Kim et...with polylinkers for the addition of gene-specific 1 kb homology arms flanking a FLOXed neoR gene. pAAV-SEPT-Acceptor...since it routinely generates knockin efficiencies of 1-40% with G418-selected clones. However, there are ...efficiency of gene editing is occasionally less than 1%, and virtually never exceeds 40%. While this is comparable...gene inactivation in human cancer.     References 1. Russell, David W., and Roll K. Hirata. "Human gene...human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor ...

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

    Type
    Blog Post
    ...minutes, which was fun to watch. A simpler 1 protein - 1 RNA system Drs. Dounda and Charpentier met at...to direct cleavage the result was a simpler 1 protein - 1 RNA system.  Dr. Charpentier advocated for the...with genes knocked out in the insulin signaling cascades in only 4 months compared to a year or greater...

  10. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...modifications). Ready to get started?   Figure 1: Break free from the ball and chain!   Epigenetics...?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental to...doi.org/10.7554/eLife.34870.  Kaas GA, Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H,...Recently, however, a particular tool has gained widespread popularity – and notoriety – as a tool of choice...leave you with a few practical considerations: Design your gRNA to overlap the CpG of interest. Having..., too), but if this is not possible due to gRNA design restrictions, anything within 5 base pairs should...

  11. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4...detects samples at clinically relevant concentrations (1-3 fM; 1x106-fold lower than before), and our combined... health, energy, and sustainability.   References 1. Pardee, Keith, et al. "Rapid, Low-Cost Detection ...Specifically this includes primer directed isothermal RNA amplification, new toehold switch designs, CRISPR/Cas9...is an Assistant professor at the Biodesign Center for Molecular Design and Biomimetics and the School of...development of diagnostics. Mostly transmitted by Aedes mosquitoes (aegypti and albopictus) and, aided by... toehold switches, which allow for the rational design of RNA sensors that can detect virtually any RNA...

  12. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...while its nucleolytic activity is eliminated (Fig. 1) (1, 3). The gRNA can be further modified by incorporation...introduce a desired genetic alteration. Listen to Our Podcast Interview with Alex Chavez Figure 1: Schematic...which Cas9 protein is directed to which target site (1). Although this is possible, this strategy suffers...sequence or regulating gene expression.     References 1. Esvelt, Kevin M., et al. "Orthogonal Cas9 proteins...genetic locus when complexed with an appropriately designed small RNA, or guide RNA (gRNA). The gRNA conventionally... truncated such that it exhibits 15 or fewer nucleotides of complementarity to a target site, Cas9’s DNA...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased...

  13. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...them to target particular cell types. Figure 1: Diagram of AAV components. Created with BioRender.com...depends on your choice of promoter/enhancer. Table 1: AAV capsids and receptors. Capsid Primary receptor...required)2 Human fibroblast growth factor receptor 1 (FGFR1)3 αVβ5 and α5β1 integrins3 Hepatocyte growth...structure of AAV2, so they are formally called rAAV2/1, rAAV2/2, and so on, although we often use abbreviated...utilizing various AAV serotypes. Scientific Reports, 15(1), 20927. https://doi.org/10.1038/s41598-025-05518-...-directed transduction. Nature Communications, 14(1), 3792. https://doi.org/10.1038/s41467-023-39554-7... and Transduction by Adeno-Associated Virus Types 1 and 6. Journal of Virology, 80(18), 9093–9103. https...

  14. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    ...been characterized. See Box 1 in Kudo et al. for more details on the design of the bNLS and NES sequences...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...using translocation reporters. Nature protocols, 13 1, 155-169. PMID: 29266096. Find the KTR plasmids here...reporters are challenging to design and difficult to multiplex. The Covert Lab provides an alternative tool with...fluorophores can be multiplexed together. Challenging  to Design: FRET reporter activity depends on both the distance...advantages. First, they are easy to multiplex. Their design allows flexible swapping of fluorescent proteins...overlap are used together. Second, KTRs are easy to design. The key determinants of KTR function are the positions...

  15. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...the genome, which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and NgAgo genome engineering...it was first published in Nat Biotech in May 2016 (1), NgAgo has received much attention and the plasmid...Pooran's NgAgo Survey Survey results As of August 1, 2016, a total of 165 researchers responded to the...the cell, the same method used by Gao et al, 2016 (1). View the Complete and Up-to-Date Results of Pooran's... dreams of a sunny day in Edinburgh.   References 1. Gao, Feng, et al. "DNA-guided genome editing using...researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid to edit a gene ...Try NgAgo for Yourself On the other hand, the widespread success of CRISPR-Cas9 across multiple model ...

  16. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...range of titers. We usually dilute our samples 1:6 million to 1:25 million.  After making the dilutions, they...note that the dynamic range of the ddPCR is between 1 and 100,000 genome copies (GC) per reaction. Since...In the image to the right, there is approximately 1 positive copy per microliter in the no template control...elongation time to 2 minutes and the denaturation time to 1 minute has been shown to increase droplet separation...diagnosis." Emerging microbes & infections 7.1 (2018): 1-9. PubMed PMID: 29691363. PubMed Central PMCID: PMC5915492... to a new plate containing the mastermix which includes primers and a ddPCR supermix. Note that the supermix...

  17. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...undergraduate students (1, 2), but educators face significant challenges in designing suitable projects. In...6). Methionine synthesis in S. cerevisiae (Figure 1) occurs through a well-characterized pathway with ...sequences for ten different MET/Met genes (Figure 1) from the two yeast species into the pYES2.1/V5-His...selected from different parts of the Met pathway (Fig. 1). In addition to the complementation experiments that...about Clare at the Biology Dept. site.  References 1. American Association for the Advancement of Science...Over Time project The Pathways over Time project provides a framework for these undergraduate research questions...

  18. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...very helpful to confirm primer design.  Use minimal template DNA Use 1 ng of PCR template DNA to minimize...two-step, 2 hr protocol prior to transformation (Figure 1). As this approach requires no special reagents or...with linearized vector backbone. DpnI digestion - 1 μl DpnI is added to the PCR mix to remove parental... by PCR, so we usually add DMSO (3%) and Betaine (1 M) to the PCR mix. When PCR is not possible Some plasmids...Twitter @jakefwatson and @JGarciaNafria.  References 1. Jones, Douglas H., and Bruce H. Howard. "A rapid ...ultracompetent (109 CFU/μg) cells. Primer design Using specific primer design (Figure 2), IVA cloning can be used...primers that bind at either side of the undesired portion, designed to amplify the rest of the vector sequence...

  19. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...cellular proteins are modified through phosphorylation (1). However, the expansion of studies on protein phosphorylation...three components required for translation (Figure 1): A tRNA that recognizes a specific ‘open’ codon ... open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...modification allowed the deletion of release factor 1, the protein responsible for terminating translation...Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing... can follow her on Twitter @nieminm.   References 1. Cohen, Philip. "The origins of protein phosphorylation...site-specific incorporation of phosphoserine (as described below) provides researchers with a valuable tool for understanding...

  20. Design Tips for Prime Editing

    Type
    Blog Post
    ...transcriptase (RT) template including the desired edit. Figure 1: Prime Editing schematic. Created with...great tips to guide your experimental design, especially the design of the RNA sequences you’ll use in prime...principles pegRNA design If you’re just starting out with prime editing, you’ll need to design and optimize... be edited and the sequence of the edit, so its design changes for every new prime editing application...transcriptase template, starting with about 10–16 nucleotides. For longer templates, it is even more important...and PE5b: In this approach, the nicking sgRNA is designed to bind only after the edit is installed. This...approach. Figure 7: Nicking sgRNAs can be designed to target edited DNA, so that nicking only occurs...
Showing: 161 - 180 of 704 results