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We narrowed to 930 results for: Gene

Showing: 441 - 460 of 930 results

  1. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many ...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520... exhaustive refinement, it was not possible to generate an optimal Rluc8-Gα construct for two of the 16...s0014-5793(02)02775-8 Additional resources on the Addgene blog Browse all of our fluorescent protein blog...Find other fluorescent biosensors Resources on Addgene.org Browse the catalog of fluorescent protein plasmids...

  2. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ...sequence of a thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968. 7. ...nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989): 245-254. PubMed PMID: 2753355. 12. ...blood-borne bacterial pathogens by use of a novel 16S rRNA gene PCR-ligase detection reaction-capillary electrophoresis...restriction cloning), adaptor ligation for next generation sequencing (NGS) library construction, and circularization...fragments end-to-end without the exo/polymerase generation of annealed complementary regions. Why use a... does fidelity mean for a ligase? DNA ligases generally prefer fully Watson-Crick base-paired dsDNA substrates...as a correct base pair. DNA ligases have been generally found to have a higher discrimination at the upstream...

  3. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...horizontal gene transfer, which occurs when bacteria of different evolutionary lineages share genes that can...bacteria that occurs when bacterial cells regulate gene expression in response to changes in cell density... the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the General Meeting ... were very valuable when shared with the next generation of scientists. The connection between mentors... with experience mentoring in STEM, including Addgene’s Joanne Kamens. Expanding Binning Singletons to...Binning Singletons, Inc. Additional resources on the Addgene blog Browse our blog posts on scientific conferences...

  4. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...description of the VGS process and use cases in Human Gene Therapy. Existing methods for AAV quality control...Adeno-Associated Viral Genomes in Academic Settings. Human Gene Therapy Methods 30:195–205 . https://doi.org/10.1089...and tips Resources on Addgene.org Find AAVs at Addgene Learn more about Addgene's viral service Find protocols...we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform that allows us to...methods, we showed that the DNA extract is a heterogeneous mix of single-stranded and double-stranded species...interest.   While the VGS process isn’t new to Addgene, we are excited to share the procedures and Python...Vector Preparations by Single-stranded Virus Next-generation Sequencing. Molecular Therapy - Nucleic Acids...

  5. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ... vector that is better suited for analyzing the gene’s function. The process is shown graphically in the...figure, in which we add EcoRI and NotI sites to Your Gene of Interest (YGOI) for ligation into a recipient..., you should use a DNA analysis tool, such as Addgene’s Sequence Analyzer, to allow you to identify which...restriction site to improve cutting efficiency. You can generally add any 6 bases, but you should ensure that the...with 18bp of homology to the ORF). We now need to generate the reverse-complement of this sequence so that...that we can successfully amplify the ORF. You can generate the reverse-complement using existing software...the final product. Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch...

  6. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...also allows for screening scFv generated from the B cell antibody genes from diseased or vaccinated humans...   Some phage display libraries screen antibody genes from B cells of immunized animals, but it's possible....1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment of HIV ...also allows for generations of scFvs against two antigens that are impossible to generate scFvs against ...probe, but is easier and cheaper to generate since it’s genetically encoded.   Translational applications... of a tagged protein. Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...-linker-VH and VH-linker-VL configurations can generate functional scFvs; however, some individual scFvs...

  7. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...amplification kit. In this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also contain...amplification by primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds...like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID-19 testing ...protocol for Cas13-based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA ...of temperatures varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources Many thanks ...Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material. Cold Spring Harbor Laboratory. https:...20detection%20(updated).pdf Additional resources on the Addgene blog Read more about SHERLOCK and DETECTR Learn...

  8. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    Published
    Sept. 16, 2025, 1:15 p.m.
    ...successfully treated with a CRISPR gene editing therapy personalized to his genetic mutation. “Baby KJ” was diagnosed...(2025). Patient-Specific In Vivo Gene Editing to Treat a Rare Genetic Disease. New England Journal of ...born with a C→T mutation on one allele of his CPS1 gene, which could be repaired with an A→G edit on the...administered the first dose of his personalized genetic medicine (Musunuru et al., 2025). Much has been...collaborative scientific effort. As we like to say at Addgene: sharing speeds science. Here, we’d like to highlight...in cellular models of KJ’s CPS1 mutation. Every genetic target is slightly different, so more options mean...Kleinstiver’s lab engineered new variants of S. pyogenes Cas9. While the wild-type enzyme requires a PAM...

  9. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ...Gibson assembly, imagine that you want to insert your gene of interest into a vector with a large tag at the...by using Gibson assembly, you can insert both the gene of interest and the tag sequences into the vector...CATCH enables one-step targeted cloning of large gene clusters. Nat Commun. 2015; Sep 1;6:8101. PubMed...the J. Craig Venter Institute in 2009. Here at Addgene we added this option to our drop down menu of common...which chews back the 5’ ends of the fragment, generating long overhangs which allows the single stranded...advantage is that this process makes it easy to generate wild type and mutant constructs at the same time...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...

  10. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...may be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable...important ways to spatially and temporally control gene expression, especially in in vivo models, and new...at least six generations. In addition to the neuronal pThy1-Brainbow constructs, Addgene also has two ...Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, 2.1 plasmids...coupling of Cre-lox with FLP-FRT recombination to generate additional recombination events, and the synthesis...and sea anemone mKate2.) They then successfully generated custom antibodies to each of these proteins and...analysis. To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins...

  11. Hot Plasmids - January 2023

    Type
    Blog Post
    Published
    Jan. 31, 2023, 2:15 p.m.
    ...were compared head-to-head for gene expression changes of non-target genes and RNA-seq of a nuclear (MALAT1...among the first batch of antibodies we produced at Addgene and, as you might suspect, it has been one of our... need a second GFP antibody?  Similarities:  Generated against full length GFP and converted to recombinant...always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid 18662... [N86/8R] page for more details. Image credit: Addgene.   Find anti-GFP [N86/8R] here!   Andrews NP...

  12. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ... provide a highly versatile means of delivering genes to many different cell types with minimal immunological...effects. AAVs are used in experiments targeting genes to specific cell types in the brain, mapping neuronal...Resources at Addgene.org Addgene's Viral Service Find Retrograde AAV Browse Optogenetics Plasmids ... to our viral vector pages on the Addgene website and on the Addgene blog to get some ideas for future...Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering Primer on Optogenetics Resources... Addgene recently attended the Society for Neuroscience 2017 conference in Washington DC (#SfN17). This...newbies might want to get into the field. Find Optogenetics Plasmids for Use in Neuroscience Research Neuroscience...

  13. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced...lentiviral vectors can be used for gene delivery Resources on Addgene.org Visit our Stem Cell Pages Find...from the patient in vitro then correct the damaged gene in the resulting iPSCs before differentiating the...PMC3987909. 6. Ban, H., et al., Efficient generation of transgene-free human induced pluripotent stem cells...Mukherjee. The ultimate goal in the field of regenerative medicine is to replace lost or damaged cells...cell is converted to a different cell type for regeneration and repair and situations where one process ...plasmids used for these methods can be found on Addgene’s stem cell page. On this page, you can also find...

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

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...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...

  15. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...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...

  17. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Oct. 29, 2019, 2:52 p.m.
    ...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...

  20. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...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...
Showing: 441 - 460 of 930 results