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We narrowed to 18 results for: REP

Showing: 1 - 18 of 18 results

  1. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...reading frames, Rep and Cap, flanked by two 145 bp inverted terminal repeats (ITRs). The Rep region encodes... Terminal Repeat, contains a Rep binding element (RBE) that serves as a binding site for Rep proteins ...plasmid Rep in trans The rep gene encodes four proteins that are required for viral genome replication and...Packaging plasmid (also known as Rep/Cap plasmid) — containing the Rep and Cap regions necessary for the...integration is mediated by proteins Rep78 and Rep68, which can bind to specific Rep-binding elements present in...DNA replication and packaging. 3' ITR (RITR) in cis Right Inverted Terminal Repeat, contains a Rep binding...Recombinant AAV constructs are designed by replacing the Rep and Cap regions between the two ITRs with ...

  2. Chemogenetics Guide

    Type
    Guide
    ...optogenetic inhibition. Sci Rep, 5 , 14366. https://doi.org/10.1038/srep14366 PMID: 26399324 Vardy, E., ...control expression of a chemogenetic reporter ensures that the reporter remains silent until a cell or tissue-specific...currently one Gs-DREADD, rM3D, that was created by replacing the corresponding intracellular region of a turkey...neurons through manipulation of ion channels arePharmacologically Selective Actuator Modules (PSAMs, pronounced...out Addgene's collection of ready-to-use AAV preparations of chemogenetic plasmids . Additionally, genetically...

  3. Optogenetics Guide

    Type
    Guide
    ...neuronal activity in vivo. Sci Rep, 5 , 14807. https://doi.org/10.1038/srep14807 PMID: 26443033 Wietek, J.,...you need. Looking for viral preps? We also offer ready-to-use AAV preparations of many optogenetics plasmids...of proteins) and sensors (genetically-encoded reporters of molecular signals). These areas have each since...

  4. CRISPR Guide

    Type
    Guide
    ...sequence. The resulting DSB is then repaired by one of two general repair pathways: the efficient but error-prone...directed repair (HDR) pathway. HDR relies on copying DNA from a matching template to accurately repair or fill... NHEJ is a naturally occurring repair pathway and is the primary repair mechanism for DSBs. While this...base excision repair pathway, which fully removes the uridine from the DNA and replaces it with another... tool to activate, repress, visualize, and isolate genes. Activation or Repression of Target Genes Targeting...sufficient to repress transcription by blocking initiation. More robust transcriptional repression (CRISPR ...demethylation DNMT3A — repression through cytosine methylation MQ1 — repression through cytosine methylation...

  5. Molecular Biology Reference

    Type
    Guide
    ...Origin of Replication ( ori ) DNA sequence which directs the initiation of plasmid replication in bacteria...advantage of the process of DNA replication in vitro. During replication, the DNA helix is unwound by a...same nucleotide bases as DNA, except thymine is replaced by uracil (U). Not all regions of DNA code for...technology has overcome this obstacle by enabling the preparation of more manageable DNA fragments. In 1952, Joshua...that typically carry genetic information and can replicate independently from chromosomal DNA. Although they...conjugation. Like chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter cell receives...genetic element that you are interested in. Self-replicate — Once you have constructed a plasmid, you can...

  6. Adenovirus Guide

    Type
    Guide
    ... DNA replication and host cell manipulation. Late transcripts are expressed after DNA replication starts...production of replication-competent adenoviruses (RCAs) that would be able to infect cells and replicate autonomously...Terminal Repeat, serve as self-priming structures that promote primase-independent DNA replication. 3' ITR...Terminal Repeat, serve as self-priming structures that promote primase-independent DNA replication. Homology...adenoviral vector that has the ability to replicate and reproduce within host cells as a result of a spontaneous...heavily-spliced transcripts flanked by two inverted terminal repeats (ITRs). Adenoviral genes are divided into early...were discovered as a contaminant of adenovirus preparations, which is how they got their name, but they ...

  7. Lentiviral Vector Guide

    Type
    Guide
    ...tropism. Replication-competent The ability of a virus (or viral vector) to replicate and reproduce within...Plasmids Collection In-Stock Lentiviral Preps Addgene's Viral Prep Service Viral Vectors 101 eBook Addgene's...hosts in order to continue their lifecycle and replicate. This feature can be harnessed for stable delivery... genes, accessory genes, and the long-terminal repeats (LTRs) necessary for integration. Not all of these...of safety improvements and the potential for replication-competent lentivirus production — that is, creation...particles that could infect cells and further replicate on their own. While the viral components are still...Interchangeable; usually encodes for VSV-G Safety Safe; replication incompetent by using three separate plasmids ...

  8. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...tropism. Replication-competent The ability of a virus (or viral vector) to replicate and reproduce within...self-inactivating viral vectors. R in cis Repeat region with repeated sequence. U5 in cis Unique 5'; in some...hosts in order to continue their lifecycle and replicate. This feature can be harnessed for stable delivery...contains packaging genes and the long-terminal repeats (LTRs) necessary for integration. Wildtype gamma-retroviruses... for safety to reduce the chance of creating replication-competent retroviruses. For a summary of all ...retroviral plasmids. This will ensure that the repeats will be maintained and often results in a greater...the only type of selectable marker. Fluorescent reporters, such as GFP, are another common selection marker...

  9. Plan Your Experiment

    Type
    Guide
    ...genome editing. Scientific Reports , 5 , 11315. https://doi.org/10.1038/srep11315 PMID: 26066060 Zukher, ...experimentally verified Cut Nick Homology Directed Repair (HDR) Generate specific genomic edits, like point...fusions with proteins that promote HDR Requires a repair template Lower editing efficiency compared to NHEJ...pegRNA optimization can be challenging Prime Edit Repress or Interfere (CRISPRi) Reduce gene expression without...sequence dCas9 or dCas9 fusion with transcriptional repressor (such as dCas9-KRAB) dCas9-KRAB can be more effective...requires donor DNA to serve as template for the repair. Once you settle on your gRNA, you can design a...target in your samples to ensure there are no discrepancies with the reference sequence you used for gRNA...

  10. Promoters

    Type
    Guide
    ...either negative or positive. Repressor proteins bind to negative-repressible operons and prevent transcription...operon is normally transcribed until a repressor/corepressor binds to the operator preventing transcription...polymerase), an operator (a segment of DNA in which a repressor or activator can bind), and the structural genes...transcription. When an inducer molecule binds to the repressor, it changes its conformation, preventing its binding...negatively-controlled operon (Figure 3). A positive-repressible operon works in the opposite way. The operon ...Lac operon; constitutive in the absense of lac repressor (lacI or lacIq). Can be induced by IPTG or lactose...Promoter of the arabinose metabolic operon trp Repressible by tryptophan Promoter from E. coli tryptophan...

  11. Guide to Using Pooled Libraries

    Type
    Guide
    ...Amplification Types of Screens Resources Pooled libraries represent a powerful tool for forward genetic screening,... simplest level, pooled libraries are single preparations of many different plasmids. Plasmids within ...either case, next-generation sequencing of the maxiprep DNA is recommended to verify that the library ...experiments, and can also negatively affect data reproducibility. Figure 1: Simplified flow chart for amplifying...To make sure that every plasmid is adequately represented in the population, you’ll need to infect many...of each cell carrying it as being a biological replicate for that plasmid. Library screens can be divided...process: Amplify the library (electroporation and maxiprep). If delivering as virus, make virus; this creates...

  12. Molecular Cloning Techniques

    Type
    Guide
    ...directly into E. coli , where the normal replication process will repair the nicks. It is important to note ...interest and then insert them into plasmids for replication and protein expression. There are many methods...note that restriction enzyme target sites can be repeated throughout a specific DNA sequence. This can make... circular product with four nicks that bacteria repair after transformation. LIC does not require site-specific... has difficulty assembling DNA fragments with repetitive sequences and DNA that ends in sequences that...more bases of homologous ends, the endogenous DNA repair pathways will produce the fused product. One major...

  13. Modular Cloning Guide

    Type
    Guide
    ...constructs with custom repeat arrays, containing anywhere between 12 and 31 repeats. Golden Gate TALEN Accessory...Plasmids with control elements and fluorescent reporters to be used in combination with the MoClo-YTK toolkit...multi-part assemblies in either integrative or self-replicating plasmid vectors for working in cyanobacteria....Cultivarium Tools 189 plasmids, including origins of replication and selection markers for a variety of bacterial...

  14. Antibody Guide

    Type
    Guide
    ... engineered small proteins derived from ankyrin repeat proteins. These are antibody mimics instead of ...isotype. Overview of application Fix your samples. Prepare your samples using permeabilization, antigen retrieval...and controls follow that of IF (above). Sample preparation, signaling excitation, and assay readout vary...must also be run. Unique samples (non-biological replicates) often each require their own set of controls...conjugating them in the lab? How will the sample be prepped, and will this or other factors affect epitope ...

  15. Educational Resources

    Type
    Guide
    ... How-to Blog Read about plasmids in Addgene’s repository , and related molecular biology techniques. Topics...

  16. Science Guides

    Type
    Guide
    ...Pooled Libraries Pooled libraries are single preparations of many different plasmids. Plasmids within ...

  18. Sequencing Primers

    Type
    Guide
    ...CCCGAAAAGTGCCACCTG 5' of MCS in pQE vectors Forward pREP Forward GCTCGATACAATAAACGCC Rous sarcoma virus (...
Showing: 1 - 18 of 18 results