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We narrowed to 1,141 results for: Gene

Showing: 621 - 640 of 1141 results

  1. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...enzyme cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between these...Schematic of SLIC. To start the SLIC cloning process, a gene of interest is PCR amplified to add 5′ and 3′ homology... LIC received an important update, courtesy of Addgene depositor Stephen Elledge. His new method, named...flexible. For one thing, those 5′ overhangs can be generated in several ways. One option, imprecise T4 digestion...work directly on PCR products, with 5′ overhangs generated either by annealing a mixture of products or from...regions from the target vector. 5′ overhangs can be generated by T4 polymerase (left branch); by annealing two...concentrations of insert to succeed. 5′ overhangs are generated in the linearized vector by T4 digestion; then...

  2. All in a Twist: dsRNA

    Type
    Blog Post
    ...dissociate when gene transcription is complete. However, transcription of thousands of genes is constantly...store their genetic information. All of these viruses have one thing in common – they can generate dsRNA as... Here, we will cover how these structures are generated, why it’s important for cells to sense their formation...inherently double stranded, while ssRNA viruses can generate dsRNA intermediates as part of the viral replication...as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription or as inverted...inverted repeats. These elements are almost always epigenetically silenced, but dysregulation of these systems... ionizing radiation, can also result in dsRNA generation. While these molecules are clearly associated...

  3. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...observed in mammalian cells is to tag an endogenous gene. By imaging cells (or tissues) that produce an endogenous...fluorescent proteins or using endogenous tagged genes) can be used to verify performance under different... develops genetically encoded FRET based biosensors.     Additional Resources on the Addgene Blog Which...mVenus as a reference protein is available from Addgene. Another way to dodge cellular variation observed...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him... Protein Biosensors Additional Resources on Addgene.org Find Empty Backbones with Fluorescent Tags Find...

  4. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...compatible approach to simultaneously clone two genes-of-interest (GOI) into two independent expression...from a different plasmid, which may lead to unequal gene dosage and high variability of protein coexpression...    Listen to this episode of Hot Plasmids! Optogenetic control of CRISPR-Cas9 by CASANOVA Article contributed...type II CRISPR systems including the popular S. pyogenes Cas9 (Rauch et al., 2017, Hynes et al., 2017)....biological tool, Niopek and colleagues turned to optogenetics. Through the use of light-sensitive protein ... “CRISPR-Cas9 activity switching via a novel optogenetic variant of AcrIIA4.” CASANOVA responds to blue...What’s great about CASANOVA? It works with the S. pyogenes Cas9, dCas9-effector fusions, and xCas9, and requires...

  5. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    ...ion-pump encoding genes HCN1 KCNJ2 KCNJ13 GABRG1 KCNIP2 KCNQ1 Examples of some genes encoding gap junction...resources on the Addgene blog: Addgene Optogenetics guide Primer on Optogenetics blog series Addgene Biosensors... light-based genetic tools for studying bioelectricity in vivo. Here are some optogenetic and GEVI tools... Aurora and Phobos optogenetic tools - A collection of bi-stable anion optogenetic channels which can ... al. "Use of genetically encoded, light-gated ion translocators to control tumorigenesis." Oncotarget ...ion-channels and pumps in developmental biology, regeneration studies, and cancer biology. I was first introduced...affect patterning and in some cases even override genetically encoded patterning. Based upon trends in studies...

  6. Hot Plasmids - May 2022

    Type
    Blog Post
    ...dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system...Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang, et...reduces the need to generate “single-use” transgenic lines. Instead, researchers can generate and maintain transgenic...each protein of interest and, for in vivo work, generate a dedicated transgenic line. Using zebrafish as... the liver.    Fig. 2: Summary of the generational evolution of the AAV capsids (left) and transduction...

  7. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...editing, this epigenetic strategy doesn’t induce DNA damage or permanently alter a gene product; it simply...on Prime-editing-Assisted Site-Specific Integrase Gene Editing (PASSIGE), their previously-reported technique...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...optimizations for prime editing CHARM: A compact epigenetic silencer Recombinant antibodies for marking astrocytes...magnetic beads for scarce macromolecules in heterogeneous samples. bioRxiv 2024.01.21.576499; doi: https...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that...with ribonucleoprotein cargo. eVLPs contain no genetic material, which ensures the editor will only be...

  8. The 12 Days of CRISPR

    Type
    Blog Post
    ...cells🎵 On the 3rd day of CRISPR, Addgene gave to me: dCas9-based gene editing for cleavage-free genomic... of CRISPR, Addgene gave to me: Cas-E-le-ven! (Cas11)🎵 On the 10th day of CRISPR, Addgene gave to me:... day of CRISPR, Addgene gave to me: RADAR from Gao🎵On the 8th day of CRISPR, Addgene gave to me: Functional...CRISPR, Addgene gave to me: updated CRISPR101 blog posts!🎵 On the 4th day of CRISPR, Addgene gave to ...let's get started!  On the 12th day of CRISPR, Addgene gave to me: Noninvasive assessment of gut function...Functional Genetic Variants Revealed by Massively Parallel Precise Genome Editing (CRISPEY!)🎵 On the 7th...7th day of CRISPR, Addgene gave to me: Drag-and-drop genome insertion, without DNA cleavage, with CRISPR-directed...

  9. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...that there are approximately 950 different human genes encoding GPCRs (Takeda et al., 2002). G proteins...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520... on Addgene.org Check out Addgene's chemogenetics guide Find chemogenetics plasmids for your research...cell type. Chemogenetics is similar to optogenetics but instead of light, these genetically modified receptors...neurons and specific behaviors. Chemogenetics uses genetically engineered receptors to interact with small...are ideal candidates for use in chemogenetics. To create chemogenetics receptors, GPCRs were first mutated... about chemogenetics and a concise table of DREADDS and their activity in neurons on Addgene’s chemogenetics...

  10. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...Alkaline phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally...GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes that affect autophagy (ATG) in yeast. Atg8p (“p”...fluorescent biosensor that was recently deposited with Addgene by Dr. Mark Prescott. This system was developed...via the secretory pathway, and then cleaved to generate an active form of the protein. To monitor autophagy...Pho8Δ60 in the vacuole, where it is cleaved to generate an active enzyme. Pho8 phosphatase activity or.... cytosol, mitochondria), rather than tracking generic autophagy markers. It’s particularly useful for...mitochondria-targeted variants are available from Addgene. References 1. Rosado, C., Mijaljica, D., Hatzinisiriou...

  11. The Codon Guide to Synthetic Biology

    Type
    Blog Post
    ...studies for various categories, including CRISPR gene-editing, the foundations of synthetic biology, DNA... the editor of Asimov Press and is designing a genetic engineering curriculum at MIT. He also writes Codon...

  12. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...triggered by engineered nucleases." G3: Genes| Genomes| Genetics 5.3 (2015): 407-415. PubMed PMID: 25566793...off-target mutation and not an edit in the target gene. The validation process, also known as CRISPR genotyping.... 12. Osborn, Mark J., et al. "Evaluation of TCR gene editing achieved by TALENs, CRISPR/Cas9, and megaTAL...as the field seeks to generate reproducible, quality data on the role of genetic networks in biological... the Addgene Blog Learn to Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate Mouse... CRISPR experiments, many are turning to next-generation sequencing options over older assays. Biased... therapy. Low frequency off-target effects may generate irreproducible data in a research setting, but...

  13. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...for stable fly cell lines; Hygromycin resistance gene; for genome integration by spontaneous insertion...own Fragmid components! Ensure your backbone and genes contain only the Bbsl cut sites needed for the Fragmid...plasmids on the Addgene website. Step 4: Naming your assembled plasmid and generating the vector file....2024.100519 More resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: ...been looking for just the right CRISPR vectors on Addgene and found instead ones that were… pretty close,...select the components they want for each module, generate the vector file, and quickly clone their vector...Fragmid components are available for request on the Addgene website, both as individual plasmids and as the...

  14. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...you attempt to verify the starting methionine of a gene but the blastx results start the alignment at a ...unlikely to have an exact entry for your favorite gene fused to an epitope tag or fusion protein. If your... our Snapgene-powered plasmid maps. Resources on Addgene.org Sequence Analysis of your Addgene Plasmid...This post was updated on Dec 4, 2017. At Addgene, we continually use the Basic Local Alignment Search...process has changed with the introduction of next generation sequencing!  Choosing a BLAST program Of the...databases, and elements of the BLAST search pages. At Addgene, we use blastn to identify any discrepancies in... viewing blastx results. When using blastx at Addgene, we use the default “Non-redundant protein sequences...

  15. 8 Great Depositor Research Stories

    Type
    Blog Post
    ...tool for large inserts.  DAP arrays for multiplex gene editing: Guest blogger Qichen Yuan tells us about... share on our blog, please reach out to news@addgene.org - we'd love to hear from you!       ...

  17. Recombinases AAV Preps

    Type
    Collection
    ...site-specific recombinases, which can be used to control gene expression. See our Cre-lox Plasmids page for more...

  18. Adenovirus Plasmids

    Type
    Collection
    ...adenoviral plasmids. Want an adenoviral plasmid with your gene of interest? Search our database for adenoviral ...capacity (>8 kb), high titers, and high levels of transgene expression. Additionally, these viruses are able...Disadvantages of adenoviruses include transient transgene expression, as well as the substantial immune ...transfected into mammalian packaging cell lines to generate adenovirus particles. Read our Adenoviral Guide...terms Adenoviral Strains Bacterial strains for generating adenovirus. ID Strain Description PI 16398 BJ5183... Shuttle For production of viruses containing transgene under a chosen promoter Vogelstein 16403 pShuttle-CMV... Shuttle For production of viruses containing transgene under CMV promoter Vogelstein 16404 pAdTrack Shuttle...

  19. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced...induces cytoskeletal disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal...Also, avoid unnecessary talking because talking generates microbe-laden aerosols that may enter the hood...many times and could be contaminated.Learn how Addgene uses barcodes to track samples Keep track of passage...Second  Edition), 2012. Additional Resources on the Addgene Blog The Challenges of Cell Culture Cloning Mammalian...Plasmids 101: Mammalian Vectors Resources on Addgene.org Find mammalian plasmids for your research Find...

  20. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...17179932. Additional Resources on the Addgene Blog AAVs CREATed for Gene Delivery to the CNS and PNS Advanced...Neuroscientist's View Control Gene Expression with FLEx Vectors Resources on Addgene.org AAV Vector for TVA Expression...selectively infect a genetically defined cell population. While the EnvA genetic sequences are supplied...combination with molecular tools (e.g., optogenetic or chemogenetic receptors) enables complex control of...enter cells and deliver a variety of cargoes, genetically engineered neurotropic viruses emerged as powerful...individual neurons they infect. However, they generally are not transmitted to adjacent neurons via synapses...of the fact that RABV is an enveloped virus. In general, viral envelopes dictate infection of target cells...
Showing: 621 - 640 of 1141 results