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We narrowed to 956 results for: REP

Showing: 381 - 400 of 956 results

  1. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...sequences required for viral replication and transcription as well as reporter genes. To learn how you can...include activation and repression of your target gene, activation and repression of different genes in ...Lab at MIT developed plasmid based systems that report the order in which specific chemical inputs are...recombinase-based state machines could potentially be used to report upon and/or engineer “temporally ordered biochemical...derived from Zika virus and, upon binding, produce reporter proteins. These toehold switches can be used to...

  2. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    ...as the repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template...new era in C. elegans research. Homology directed repair (HDR), which enables insertion of custom genomic... a fluorescent protein fusion, transcriptional reporter, and loss-of-function allele in just one injection... CRISPR/Cas9 applications, SEC selection could replace other, more restrictive C. elegans selectable marker.... This novel, rapid, and user-friendly system represents another leap forward for the CRISPR/Cas9 revolution...Editing Efficiency CRISPR 101: Homology Directed Repair  ...

  3. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...will amplify even if its target states are only represented in a very small number of registers. Instead,...that uniquely express selectable and/or sortable reporters in certain states of interest. For example, in...above (Sanger sequencing, qPCR, and fluorescent reporter-based) each come with their advantages and disadvantages...the most convenient strategy is the fluorescent reporter based method since it lends itself to cheap, quick...detection with flow cytometry. However, expressing reporters will consume cellular resources and therefore ...The input plasmid (high copy, ColE1 origin of replication), contains large serine recombinase genes downstream...we used to drive recombinase expression may be replaced by sensors that correspond to the desired inputs...

  4. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    ...published a report on the social and ethical issues of genome editing and human reproduction, the relevant...Nuffield Council’s report is that when it became clear that this assistive reproductive technique was feasible..., the MIT Technology Review dropped a bombshell report. A scientist working in China was using CRISPR/...question of “how do we move forward” is the June 2012 report on the ethics of “Novel techniques for the prevention...established ethical issues raised around mitochondrial replacement therapies. In other words, rather than having...

  5. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...the ligation junction were represented. Sixteen separate pools were prepared, each with a different complement...fidelity. In this method, substrate pools were prepared consisting of one target (template) strand and...and the ATP dependent 9°NTM DNA Ligase are not replacements for T4 DNA Ligase in applications such as routine...routine cloning or DNA library preparation. However, when a method relies on accurate ligation of nicks... Biolabs, specializing in the enzymology of DNA repair enzymes.     References 1. Shuman, Stewart, and...et al. "Ligase-based detection of mononucleotide repeat sequences." Nucleic acids research 27.24 (1999)..., and Stewart Shuman. "Last stop on the road to repair: structure of E. coli DNA ligase bound to nicked...

  6. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... These breaks can then be repaired by homologous recombination with a repair template.  Each zinc finger...Frederick Griffith used mouse models to prove that Streptococcus pneumoniae bacteria could be transformed from... that revolves around synthetic humans called replicants which are genetically engineered by a powerful...genetic engineering as CRISPR-Cas editing soon replaced the protein-based DNA-binding domain with a single-guide...genome. These technologies rely on the cellular DNA repair machinery, which can lead to high frequency of ... Newby GA, Raguram A, Liu DR (2019) Search-and-replace genome editing without double-strand breaks or ...10.1093/nar/gkf495 Watson JD, Crick FHC (2003) Reprint: Molecular Structure of Nucleic Acids. Annals of...

  7. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...microbiome by using engineered genetic tools and reporter microbes. Let’s take a look at a few of these ...modular suite of plasmids (pGT) with various replicative origins that can be used for broad or narrow ...is a ribosome binding site (RBS) followed by a reporter gene. The hairpin blocks translation by sequestering... reveals the RBS and allows translation of the reporter gene resulting in GFP signal. With the addition... built this toolkit upon the Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial...distribution of gut bacteria without fixation or preparation, and deliver CRISPR components for gene disruption...Central PMCID: PMC5963681.  Meyer, Richard. "Replication and conjugative mobilization of broad host-range...

  8. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...able to replicate, and for a virus to cross a synapse from one cell to another, it must replicate. Modified...infects one cell (the “starter cell”). It then replicates within that cell, and nascent RABV cross synapses...infection occurs, subsequent cycles of viral replication and retrograde transmission occur, spreading ...limitations. First, these high levels of viral replication are toxic. Second, infecting subsequently higher...G-deleted RABV will infect the starter cell. RABV replication will then occur and the G protein (supplied in...order neurons, the G-deleted RABV can divide and replicate (since these processes are mediated by other viral...

  9. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ... have all of your plasmid data, we’ll send you prepaid shipping materials (i.e., a deposit kit) with instructions...kanamycin, nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline, combinations of the above, or... sufficient quantity of DNA is produced from a miniprep and “low copy” if special growth conditions are...cloning strain DH5α. For plasmids with highly repetitive sequences (prone to recombination in bacteria...For plasmids that contain the R6Kγ origin of replication, choose the Pir1 strain. If your plasmid cannot...in the format: labname_addgene_batch_upload.xls Replace ‘labname’ with the name of your PI and send the... you a deposit kit with instructions on how to prepare and ship the samples to Addgene and contact your...

  10. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...pathways are involved in RNA based surveillance of repetitive gene families and provide a way to guide inheritance...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...water. Dr. Terns gave a great introduction to the repeated and distinct aspects of the CRISPR regions of ...gene cassettes. Thse vectors are engineered to replace the drug resistantance genes in the bacteria thus... labs are using CRISPR/cas9 and more are being reported. Zhou Songyang from Baylor College of Medicine...

  11. Using a Light Microscope Protocol

    Type
    Protocol
    ...image of that object! Equipment Light Microscope Prepared Slides (or other sample that can fit on a microscope... risk damaging the lens or your sample. You can repeat steps 7 and 8 with progressively higher magnification...

  12. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    ...first scientific poster, but unsure how best to prepare. Maybe you’re a presentation veteran, but have ...experience. No matter the reason for attending, your preparation and day-of game plan will ensure you have the...experience possible. Tips for poster session presenters Prepare for success You’ve put a great deal of thought ...it’s time to expertly show off your science.  Prepare the night before: To avoid rushing out the door...poster as a visual aid, not a crutch. Even the most prepared of us can find ourselves talking to our posters... part of a poster session, but with a little preparation and a few deep breaths the day-of, you’ll handle...Strategically traverse the sea of posters If you come prepared with a list of posters of interest, then you’ve...

  13. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...chromosome. During DNA repair, there is a chance that the two breakpoints will repair without the section... a derivative of Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors...template strand left behind that the cell can use to repair. Figure 3: General overview of the Cascade-Cas3...ssDNA past the nuclease domain, Cas3 is able to repeatedly cut the DNA at an efficient rate. These cuts ... or just random mutations if the chromosome is repaired with all the original DNA. With Cas3, you don’...References Loeff, L., Brouns, S. J., & Joo, C. (2018). Repetitive DNA reeling by the Cascade-Cas3 complex in nucleotide...

  14. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...vector preparations, we have used quantitative PCR to evaluate the physical titer of our preps. We often...often repeat this assay to confirm that our results are accurate. But what if we could repeat this assay... for ddPCR set-up. Prepare the master mix in a separate area from where you prepare your sample dilutions...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...

  15. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ... an AAV vector to express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes...dopamine beta-hydroxylase (DBH, magenta) to label norepinephrine (NE) expressing neurons and GFP (green) to ... right AAV for your system or to successfully replicate published works. Of particular importance to these...experimental settings. It is a curated set of technical reports on AAV vectors from Addgene and the CLOVER Center...Center, each one submitted by researchers like you. Reports are reviewed by Addgene scientists for clarity ...performance. Yet, most of the technical data needed to replicate and build on others’ results is available not ... in Adeno-Associated Virus Serotype 9 (AAV9) Preparations. Human Gene Therapy Methods, 30(6), 214–225....

  16. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ..., Zetsche et al. hypothesized that Cpf1 might represent a functional CRISPR nuclease that could be adapted...about the same size as Cas9’s, but with the direct repeat preceding the spacer rather than following it. ... only a short stem-loop structure in the direct repeat region is necessary for cleavage of a target. Cpf1...are difficult to modify through homology-directed repair (HDR). Cpf1 also expands the number of sites that...experimentally confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing...in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing efficiency for the Cpf1 orthologs...mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches at gRNA positions...

  17. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...these proteins, as well as the viral long terminal repeats (LTRs) are different. As a result, lentiviral and...adenoviral protein E1A, which has been shown to repress expression of HIV-1 LTRs (more information is available... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...all) Addgene transfer plasmids are replication-deficient meaning they can be used to create virus that is...Which genes are deleted or modified to result in replication deficiency? What percent of the vector components... vectors are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential...

  18. New Viral Vectors - Spring 2025

    Type
    Blog Post
    ... to conduct experiments faster, skipping the preparation steps of production, purification, and titering...customizability, choosing the cargo, serotype, and prep size they need for their experiments — and we’ve...them out in record time.  However, our in-stock preps remain a popular choice thanks to the convenience...cortical interneurons. Take a look to see if these new preps might enhance your research!   Table 1: New ready-to-use...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV-CAG-emiRFP670...help, with over 100 user-submitted viral vector reports. And don't forget to use the link below to keep...

  19. New Viral Vectors - Summer 2024

    Type
    Blog Post
    ...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 20... 20 additions we've made to our viral vectors repository since March.  Addgene is now offering an AAV2...gene expression! We've also added AAV1 and AAV5 preparations of several variants of a new optogenetic tool...Yizhar lab. Addgene is now offering AAV1 and AAV5 preparations of several variants of a new optogenetic tool...New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes ...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link...

  20. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ... organisms) -  perhaps you’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project...used to study aspects of cognition. Spiralians - Crepidula atrasolea (Jonathan Henry) This black footed slipper...real bottleneck. New gif for #MolluscMonday: #Crepidula atrasolea juvenile, light microscope with polarizing...it in a fertilized egg to get edited embryos (replacing the nucleus is still very difficult). Cuscuta ...targeted gene mutations in anoles lizards or in any reptiles at all. But new success is coming with CRISPR/...full hydra and hydra have asexual and sexual reproductive life cycles. One can inject plasmid DNA in the...adult female mosquitoes. Now they are trying to replace embryo injection (hard to do, expensive, not every...
Showing: 381 - 400 of 956 results