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We narrowed to 17 results for: REP

Showing: 1 - 17 of 17 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
    ...hardware-independent optogenetic inhibition. Sci Rep. 5:14366. PMID: 26399324 (Link opens in a new window...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...available. Browse our collection of Ready-to-use AAV Preparations of Chemogenetic Plasmids ! Table 4. Common promoters...Zhang S-C (2021). Human Stem Cell-Derived Neurons Repair Circuits and Restore Neural Function. Cell Stem...

  3. Optogenetics Guide

    Type
    Guide
    ...light-induced inhibition of neuronal activity in vivo. Sci Rep. 5:14807. PMID 26443033 Wietek J, Prigge M. 2016....optical switches Sensors are genetically-encoded reporters of molecular signals; e.g., calcium indicators...

  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 Figure...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 initiation of plasmid replication (by bacteria...recombinant DNA technology, which enables the preparation of more managable (i.e., smaller) DNA fragments...double-stranded DNA that typically carry genes and can replicate independently from chromosomal DNA. Although they...the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter cell receives...manipulating genes. For instance, most plasmids are replicated in E. coli and are relatively small (∼3000 - ...sequences for this purpose, which includes a DNA replication origin, an antibiotic-resistance gene, and a ... exists extrachromosomally in E. coli , it is replicated independently and segregated to the resulting...

  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
    ...example: To activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted...region where the edit should be made HDR requires a repair template and displays reduced efficiency compared...Base editors can make a limited set of mutations. Repress or Interfere (CRISPRi) Reduce expression of a particular...without permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting...promoter is typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive...vector or separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify...CRISPR protocols from Addgene depositors JOVE Video Reprint: Protocol for genomic deletions in mammalian cell...

  10. Promoters

    Type
    Guide
    ...negative or positive. Negative repressible operons, are normally bound by a repressor protein that prevents 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...of a negatively controlled operon. A positive repressible operon works in the opposite way. The operon ...bacteriophage lac 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
    ...Introduction to Pooled Libraries 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. Types of Pooled Library Screens In pooled ...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. Cloning

    Type
    Guide
    ...E. coli , where the nicks will be repaired by the normal replication process. It is important to note ...interest and then insert them into plasmids for replication and protein expression. So how do scientists ...note that restriction enzyme target sites can be repeated throughout a specific DNA sequence, which can ...creating a circular product with four nicks that are repaired by the bacteria after transformation. LIC does... has difficulty assembling DNA fragments with repetitive sequences and DNA that ends in sequences that...

  13. 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 ...

  14. Sequencing Primers

    Type
    Guide
    ...distribute primers. For sequencing plasmids in our repository, we've chosen primers based on the plasmid backbone...Qiagen) 5' of MCS in pQE vectors, forward primer pREP Forward GCTCGATACAATAAACGCC (Invitrogen) Rous sarcoma...

  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 ...
Showing: 1 - 17 of 17 results