We narrowed to 17 results for: LAT;

...the isolation and purification of cellular components, such as DNA, but in practice this isolation is ...is only feasible for relatively short DNA molecules. To isolate a particular gene from human chromosomal...,000 base pairs) for physical isolation (purification) and manipulation. With current cloning technology... and manipulating genes. For instance, most plasmids are replicated in E. coli and are relatively small...common is the Illumina NGS platform for short-read sequencing. This is the platform Addgene primarily uses...fragments of DNA, or templates, are spatially separated. These attached DNA templates are then amplified...again, producing ~1,000 copies of each template. Each template is then replicated using the modified bases...

...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...

...system that enables researchers to manipulate the genome with relative ease. Over the past decade, researchers... Select Your Desired Genetic Manipulation Different genetic manipulations require different CRISPR components... donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires a repair template Lower editing...Does the resulting population need to be identical (clonal) or is a mixed population okay? Does the edit...modifications . Cell Population The first step of validation is determining the type of cell population you need ...you will likely need a clonal population. Additionally, clonal populations are useful for determining if...edited alleles. A clonal population is not always required, and a mixed population can be sufficient for ...

...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...

...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...

...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 ...

...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... 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 noncoding strand, as this is the strand...proteins which recruit RNA polymerase and control and regulate the transcription of DNA into mRNA. Promoter binding..., bacteria still have complex systems of gene regulation, like operons. Operons are a cluster of different...structural genes that are transcribed together. Operon regulation can be either negative or positive. Repressor...

...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. Read this guide to learn...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. ...

...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 ... integrates optics and genetic engineering to 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...

...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...

...Daniel Voytas 93 additional plasmids including regulatory elements, coding sequences, CRISPR parts GreenGate... for assembling large DNA fragments based on methylation-switching. 4G-Cloning Plasmid Kit Bacterial Expression...Expression, Mammalian Expression Stephan Gruber Platform for rapid assembly of expression vectors for multi-subunit...TALE-TFs) for genome editing and transcription modulation applications Golden Gate TALEN and TAL Effector...TALEN and TAL Effector Kit (Voytas and Bogdanove). Platinum Gate TALEN Kit Genome Engineering, TALEN Takashi...Yamamoto Plasmids for construction of highly-active Platinum TALENs for use in a wide variety of target organisms...

...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...

...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...

...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...

...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...

...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...

...you need to know about plasmids, from parts, nomenclature, and uses to common promoters and cloning techniques...