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We narrowed to 16 results for: LAT

Showing: 1 - 16 of 16 results

  1. CRISPR Guide

    Type
    Guide
    ...genomic edits by copying from a DNA donor template . The donor template contains the desired sequence and is...linear dsDNA templates work better for large edits or insertions. When designing the donor template, incorporating...modifying the methylation state of cytosines in a gene’s promoter or by inducing histone acetylation or demethylation...through increased H3K27 acetylation TET1 - activation through cytosine demethylation DNMT3A - repression ...repression through cytosine methylation MQ1 - repression through cytosine methylation with improved kinetics LSD1...detection platform. Based on this non-specific cleavage, a Cas13a-based molecular detection platform, termed...pathway. HDR relies on copying DNA from a matching template to accurately repair or fill in the missing sequence...

  2. Molecular Biology Reference

    Type
    Guide
    ...enable isolation and purification of cellular components, such as DNA, but practically this isolation is ...is only feasible for relatively short DNA molecules. In order to isolate a particular gene from human chromosomal... and manipulating genes. For instance, most plasmids are replicated in E. coli and are relatively small...20,000 basepairs) for physical isolation (purification) and manipulation. With current cloning technology...most commonly used is the Illumina NGS platform. This is the platform used by Addgene’s sequencing partner...of DNA, or templates, are spatially separated from each other. These attached DNA templates are then amplified... again producing ~1,000 copies of each template. Each template is then replicated using the modified bases...

  3. Antibody Guide

    Type
    Guide
    ...produces a colored product that accumulates over time which is relative to the amount of enzyme present...quantify relative amounts of protein. Includes: Western blot ELISA Capture - used to isolate specific ... one type of antibody. Monocultures of B cells isolated from animals immunized as described above are ... regions of the antibody to be independently manipulated. Antibodies produced this way are called recombinant... used while suspended in the original media or isolated and placed in a buffer. Antibody Structure Figure...antibody fragments (shown in Figure 2) to completely unrelated molecules. Examples include: scFv - Single-chain...instead of modified antibodies and are not closely related to antibodies. Monobodies - Monobodies are synthetic...

  4. Adenovirus Guide

    Type
    Guide
    ...necessary for viral DNA replication and host cell manipulation. Late transcripts are expressed after DNA replication...replication and host cell manipulation. L1, L2, L3, L4, L5 in trans Adenoviral late transcript regions that...are divided into early (E1A, E1B, E2, E3, E4) and late (L1, L2, L3, L4, L5) regions. Early transcripts ...also made them the most widely used viral vector platform for vaccine design against a diversity of viruses...which is how they got their name, but they are not related. AAVs require the presence of adenoviral genes ...and have been produced by replacing the early and late adenoviral transcript regions between the two ITRs... inside of the adenoviral capsid and directly correlates to the size of the DNA removed from the viral...

  5. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...hepatitis virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased...cis The polyadenylation or poly(A) signal aids in the nuclear export of RNA and RNA translation, and promotes...encodes for virus-associated RNAs involved in regulating translation and inhibiting cellular anti-viral defenses...which is how they got their name, but they are not related. AAVs require the presence of adenoviral genes ...available inside of the small AAV capsid and directly correlates to the size of the DNA removed from the viral...kidney 293T (HEK293T) cells are the most common platform to produce AAV vectors. Additional production ...cannot enter the lytic cycle, and instead establish latent infections by integrating into the host genome ...

  6. Chemogenetics Guide

    Type
    Guide
    ...previously used in human populations in Japan, making it an attractive option for translational studies, however...receptors, or ion channels targeted to specific cell populations (most often neurons) that can be turned on or...therefore allowing precise control over the cell population they are targeted to. To find chemogenetics plasmids...ligand. Ligand binding then activates G-proteins to modulate downstream signaling. GPCRs have been shown to...G-protein and activate neuronal firing through stimulating phospholipase C, which releases intracellular... and inhibit neuronal signaling by inhibiting adenylate cyclase and downstream cAMP production. There ...hM4Di, and is especially attractive for use in translational studies because it is FDA and EMA approved. ...

  7. Promoters

    Type
    Guide
    ...by various DNA regulatory sequences including enhancers, boundary elements, insulators, and silencers....RNA responsible for providing a stable template for the translation of a protein. RNA polymerase III transcribes...polymerase transcribes DNA to mRNA which is ultimately translated into a functional protein. Thus the promoter ...produced. The antisense strand is referred to as the template strand or non-coding strand as this is the strand...proteins which recruit RNA polymerase and control and regulate the transcription of DNA into mRNA. Promoter binding...promoter region stably and transcription of the template strand can initiate. The TATA box is a DNA sequence...proximal promoter which contains many primary regulatory elements. The proximal promoter is found approximately...

  8. Optogenetics Guide

    Type
    Guide
    ...silence the same neuronal population, or activate different neural populations at distinct times. Temporal... Two factors determine which neuronal population is manipulated in a given experiment: the expression ...optics and genetic engineering to measure and manipulate cells (frequently neurons) and their governing...neurons upon exposure to light through direct stimulation of ion channels. Naturally occurring channelrhodopsins... nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely ...Halorhodopsins are light-gated inward chloride pumps isolated from halobacteria. Wild-type halorhodopsin, known...High-efficiency blue-shifted channelrhodopsin from Platymonas subcordiformis 445 CoChR Channelrhodopsin from...

  9. Lentiviral Vector Guide

    Type
    Guide
    ...hepatitis virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased...single strand of RNA contains packaging genes, regulatory genes, accessory genes, and the long-terminal...Wild-type lentiviruses additionally require the regulatory genes tat and rev , along with virus-specific... — contains packaging genes gag and pol , and regulatory genes tat and rev ; separated into two plasmids...plasmid — contains entire viral genome (packaging, regulatory, and accessory genes), only envelope removed ...can more specifically dictate the target cell population and reduce cellular toxicity. Common Uses of ...(FACS) is used to sort cells expressing GFP and later expanded into a cell line. More information on generating...

  10. Plan Your Experiment

    Type
    Guide
    ... Select Your Desired Genetic Manipulation Different genetic manipulations require different CRISPR components...Select gene and genetic element to be manipulated In order to manipulate a given gene using CRISPR, you will...Plan Your Experiment Overview Select Genetic Manipulation Select Expression System Design gRNA Publications...a powerful system that enables researchers to manipulate the genome like never before. This section will...other organisms. First, consider the genetic manipulation that is necessary to model your specific disease...components. Selecting a specific genetic manipulation can be a good way to narrow down which reagents are...existing reagent to suit your needs. Genetic Manipulation Application Cas9 gRNA Additional Considerations...

  11. Guide to Using Pooled Libraries

    Type
    Guide
    ...name, pooled libraries are supplied as a mixed population of plasmids in a single tube. Pooled libraries...may lead to false positives or false negatives in later experiments, and can also negatively affect data...every plasmid is adequately represented in the population, you’ll need to infect many more cells than the...the cells die, removing their plasmids from the population, and only a small fraction survive. After the... serves as a non-selected control. These two populations are then sequenced using NGS to determine which...

  12. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...hepatitis virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased...(FACS) is used to sort cells expressing GFP and later, sorted cells are expanded into a cell line. You...best practices with regard to gamma-retroviral-related research. See Addgene’s Biosafety Resource Guide...Plasmid Elements Plasmid Type Element Delivery relative to transgene Purpose Transfer plasmid LTR in cis...

  13. Cloning

    Type
    Guide
    ... molecular cloning scientists can amplify and manipulate genes of interest and then insert them into plasmids...hundreds of available enzymes, many of which are relatively cheap. They also cut specific target sequences...and choice of restriction site. Because of the relatively long stretches of base pairing in the annealed...

  14. Educational Resources

    Type
    Guide
    ...any of our eBooks to learn about popular topics related to our collections. Titles include: Plasmids 101...Read about plasmids in Addgene’s repository , and related molecular biology techniques. Topics include: Genome...

  15. Science Guides

    Type
    Guide
    ...start learning about various scientific concepts related to our plasmid collection. Find links to the guides...optics and genetic engineering to measure and manipulate cells (frequently neurons) and their governing...

  16. Addgene's eBook Collection

    Type
    Guide
    ...you need to know about plasmids, from parts, nomenclature, and uses to common promoters and cloning techniques...
Showing: 1 - 16 of 16 results