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We narrowed to 924 results for: Gene

Showing: 401 - 420 of 924 results

  1. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ... maintain the genes they carry among organisms belonging to many different phylogenetic groups (Jain and...that the plasmid is conjugative and carries the genes necessary for transfer initiation between bacterial...plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197. https://doi.org/10.1016/0378-1119... S. (2011). Broad-host-range plasmid vectors for gene expression in bacteria. Methods in Molecular Biology...viscosus, Aerobacter aerogenes, Aeromonas hydrophila, Agrobacterium tumefaciens, Alcaligenes eutrophus, Azotobacter...salmonicida, Agrobacterium tumefaciens, A. rhizogenes, Alcaligenes eutrophus, Enterobacter sp., Erwinia amylovora... broad host-range plasmid RK2. Molecular & General Genetics: MGG, 152(3), 129–135. https://doi.org/10.1007...

  2. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...when trying to edit a gene using homology directed repair, since HDR-mediated gene editing is most efficient...formerly Cpf1), a nuclease that generates double strand breaks in target genes resulting in the formation ...always positioned correctly to target a particular gene. Furthermore, a target sequence may have high homology...plasmids from this publication at Addgene. Additional Resources on the Addgene Blog  Read more about Cpf1...sequences While PAM sequences for the commonly used S. pyogenes Cas9 (3'-NGG) are abundant throughout the human...circumvent this limitation: 1) the use of novel S. pyogenes Cas9 variants with varying PAM sequences, 2) the...Cas9 homologs derived from species other than S. pyogenes, and 3) the use of non-Cas9 enzymes. (For more...

  3. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...edits of interest than solely transfected cells. “Gene On” (GO)- a functional reporter system to identify... (Katti et al., 2020). They named this reporter “Gene On” or GO. GO works by affecting protein translation...fluorescent proteins. Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter...and enrich for base-editing events in vivo. TLDR: Gene On (GO) from the Dow lab is a flexible reporter ...base-pair mCherry insertion. Editing of these motifs generates lesions that are subject to excision and ssDNA...This edit changes the histidine to a tyrosine generating a GFP variant (GFPY66) that has a shifted emission...hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that contains an ACG start...

  4. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    Published
    July 28, 2015, 2:30 p.m.
    ...homology arm as a gBlocks® Gene Fragment from Integrated DNA technologies. Generally speaking, anyone using... an effort to track down every last one of those gene regulatory proteins and the places in the genome...together on several projects and this is one related to gene expression. One goal of the ENCODE project is to...with detailed protocols, go to www.addgene.org/crispr/tagging. Addgene: What motivated you to adapt CRISPR...tagging system, which they’ve now made available at Addgene. For more details on the plasmids that are currently...can now look at many more transcription factors. Addgene: How does the system work? Mendenhall: The system...antibodies. The results look almost identical. Addgene: What advantages does your system have in this ...

  5. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    Published
    April 22, 2015, 2:06 p.m.
    ... to turn on genes at different times. It requires more complex synthetic networks. Addgene: What hurdles...on an engineering project including hundreds of genes and regulation and resource control - everything...gives us enough hardware to control hundreds of genes. We’ve created enough regulatory elements, CRISPRs...the scale of genetic engineering, with the ultimate objective of genome design.” It’s genetic engineering...larger genetic circuits that could conceivably be as large as natural regulatory networks. Addgene: What...things are easy with CRISPR. Addgene: You’ve made many deposits to Addgene over the years. Can you describe...toolbox for taking on very large genetic engineering projects. Addgene: How easy is it to do this now? ...

  6. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    Published
    June 17, 2014, 7:57 p.m.
    ...the complexity of genetic circuits, allowing for tighter spatio-temporal control of gene expression within...world of possibilities in the engineering of complex gene networks. A comparable system for targeting specific...will make those new plasmids available through Addgene, and they hope others will do the same. “This isn...10.1186/1754-1611-8-7. PubMed.  Related Resources Addgene's Synthetic Biology Overview More Synbio Blog Posts...

  7. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...in neurons. (b) Genetic cargo can be customized to control transgene expression. Gene regulatory elements....     Figure 1: AAV toolkit for targeted gene delivery to the brain. The AAV (a) capsid, (b) cargo...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. ...LY6C1, with relatively even distribution of target gene expression (EGFP, in the depositing manuscript) ...Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature Protocols;14, 379–414...2017). Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems...Gradinaru, V. (2022). Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous...

  8. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...looks like the genetic cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate...with those particles. Since the Env, Gag, and Pol genes are not encoded in the transfer plasmid, the component...Similarly, if the viral vector carries an shRNA to a gene essential for cellular function, this would also...Resources Resources on Addgene.org Addgene viral services Addgene Biosafety Guide Addgene Viral Plasmids ...? That’s okay! In this article we will discuss general viral safety considerations and best practices ...of what is being integrated by the virus. If an oncogene overexpression construct is being delivered, that... and the same holds true for types of viruses generated in the lab. The four most common types of viruses...

  9. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...Additional Resources on the Addgene Blog Treating Muscular Dystrophy with CRISPR Gene Editing Learn to Use ...Cas9 have also been used to create mutations in genes essential to HIV-1 replication. However, this approach...modification targeting either the LTRs or essential genes. Many of these escape mutations were located near...Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing. Sci Rep. 2016 Mar 4;6:22555. PubMed PMID...Screens Additional Resources onf Addgene.org Check out Addgene’s newly updated CRISPR Guide! Find CRISPR...Activation Mediators (SAM) system available from Addgene. This system uses the dCas9-VP64 fusion commonly...others and accelerate the pace of research. We at Addgene hope to see the problem of in vivo CRISPR delivery...

  10. Microbes: Look, Listen, and Tell

    Type
    Blog Post
    Published
    Aug. 28, 2018, 12:04 p.m.
    ...ultrasound to image bacteria in vivo: acoustic reporter genes’ on our blog. Creative approaches to presenting...Bourdeau, Raymond W., et al. "Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian...way to visualize them. You can also “hear” them. Addgene depositor Dr. Mikhail Shapiro from Caltech presented...loads of inspiration for the current and the next generation of microbiologists. If you got inspired to produce...PubMed Central PMCID: PMC5920530.  Resources on the Addgene Blog Read Our Post on CRISPR Methods for Bacterial...Science Communication using Twitter  Resources on Addgene.org Check out Our Microbiology Resource Page CRISPR...bacteria Deposit Your Microbiology Plasmids with Addgene ...

  11. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 7, 2017, 1:58 p.m.
    ...new green glowing protein could be used to measure gene transcription, as well as to track protein localization... he was right! He began to study the A. victoria gene responsible for encoding GFP, and, in 1992, he reported...

  12. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    Published
    July 7, 2017, 1:32 p.m.
    ...Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from the...treat disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School...source biology in his own lab, at the Center for Regenerative Medicine, and within the biomedical research...Along these lines, Darrell recently joined the Addgene Board of Directors where he hopes to help keep ... Kotton and the Boston University Center for Regenerative Medicine. Darrell describes his career goals...

  13. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...Plasmids for endogenous gene tagging in human cells Resources on Addgene.org Browse all CRISPR plasmids...endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and dTAG-13 ...endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, dTAG-13, and...Figure 1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...47 Additional Resources on the Addgene Blog Easi-CRISPR: Generating Knock-In and Conditional Mouse Models.... To overcome this limitation, we created a generalizable tag-based strategy, which we refer to as the... scales that are not possible with traditional genetic approaches. In addition, the dTAG system is versatile...

  14. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...overexpressing a gene or protein after transfection). Future of FRET The first genetically encoded FRET biosensor...Resource page and can be used to create fusions with a gene of interest. These plasmids will be most useful ...designed to measure specific small biomolecules or gene activity are often intramolecular probes, as the...name a few.   Find Many FRET-Based Biosensors on Addgene's Biosensor Pages   Recent innovations have demonstrated...PubMed PMID: 26210398. Additional Resources on the Addgene Blog Tips for using FRET in Your Experiments Learn...FRET Based Biosensors Additional Resources on Addgene.org Find FRET Pairs Check Out Fluorescent Protein...

  15. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ... this plasmid will work for them. Species of Gene or Insert Choose from: H. sapiens (human), M. ... Cloning Method Choose from: Gateway cloning, gene synthesis, Gibson cloning, ligation-independent ... Stable strain. For plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival...information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene  Quality control... field, and contact Addgene. We may require that you send your samples to Addgene pre-transformed in this...the spreadsheet to deposit@addgene.org with the following information: Addgene account username. In order... to appear in your Addgene account under My Plasmids, we need to know your Addgene account username. Shipping...

  16. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...disease-causing alleles, and the growing need for gene editing in research, the old PAM constraints just...RNA will do. Similarly, if you want to knock out a gene, a frameshift mutation introduced anywhere within...take a bp or two upstream or downstream) for most gene targeting experiments. Between SpRY and several ...Resources on Addgene.org Addgene’s CRISPR Guide CRISPR Plasmids and Resources Resources on the Addgene blog ...biology, the answer is always it depends, but generally the efficiency of the desired edit is reduced ... Cas9s that recognize NG PAMs. The Nureki lab generated SpCas9-NG, a rationally engineered Cas9 with NG...name implies (Nishimasu et. al, 2018). xCas9 was generated by the Liu lab through phage-assisted evolution...

  17. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ... bright enough to detect endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc...Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are available...such as the dynamics of subcellular structures and gene expression. You must see the movies demonstrating...al. (2016) CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nature...when employing optogenetic regulators to initiate the event being monitored. Optogenetic regulators respond...wavelengths overlap with optogenetic initiation wavelengths, excitation of the optogenetic regulator leads to... be paired with optogenetic regulators. Many of these tools are available from Addgene and we're excited...

  18. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ...(MP6) permits quick, large-scale mutation of the gene of interest. They created a selection phage (SP)...transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all... a library PAM, it would induce transcription of gene III, permitting phage propagation and keeping that...evolution to generate and select for Cas9 mutants with PAM flexibility. In this system, the mutagenesis plasmid...this paper are available at Addgene.  Additional Resources on the Addgene Blog Learn about Engineered...another 72 hours of phage-assisted evolution, generating xCas9 2.0-2.6. This second round of evolution... Cas9 variants with activity on non-NGG PAMs, generating xCas9 3.0-3.13. To test xCas9 3.0-3.13, Hu et...

  19. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...Click on the gene names in the figure below to find plasmids containing that particular gene. ...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...FP properties. It is important to look out for general problems associated with fluorescent imaging such...constructs you create in your own work available to the Addgene community. Download our Markers of Subcellular ...PMCID: PMC4777285. Further Reading Microscopy U. Generfal Considerations when Using Fluorescent Proteins...Collection Get Tips for Using FRET Resources on Addgene.org Browse the Allen Institute for Cell Science ...

  20. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...22,000 protein coding genes in the human genome with fluorescent proteins. For each gene, we needed to design...way that prevents re-cutting but does not disrupt gene expression. (See section below.) For guide design... Zuckerberg Initiative. We hereby announce the general availability of new a tool for CRISPR scientists...analyses, but it also allows you to download any generated info for your own purposes, or delete the originals...at existing tools. As has been written here on Addgene, there has been a Cambrian-like explosion of software...that it can be used in the rank equation above. Generally, the greater this distance between the cut and...sequence inserted by HDR. Although this practice is generally advised, there is little detail published on how...
Showing: 401 - 420 of 924 results