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We narrowed to 419 results for: RAP

Showing: 41 - 60 of 419 results

  1. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ...APEX: engineered ascorbate peroxidase that can rapidly biotin label proteins A year after BioID became...  Find the APEX2 plasmids! TurboID and minturbo: Rapid and non-toxic proximity labeling  TurboID and miniTurbo...hours to label proteins and APEX2, although it can rapidly label proteins, requires H2O2 which can be toxic...more information, read the Split-BioID blog post.  RaPID: RNA-protein interaction detection So far, we’ve...that can regulate multiple biological functions. RapID (RNA-protein interaction detection) uses proximity...interactions (Ramanathan et al., 2018). To develop RapID, scientists engineered a mutant BirA* from B. subtilis... compared to the standard E. coli BirA* mutant. RapID consists of this mutant BirA* and an RNA component...

  2. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    Published
    July 17, 2018, 12:23 p.m.
    ...more hands on While teaching courses about gene therapy and genetic manipulation, I and other professors...similar types of thinking go into developing gene therapy strategies for various diseases. That’s when I ...skills to apply those concepts and understand how therapeutic strategies work. Find Addgene Blog posts about...pick the best strategy and propose a final gene therapy approach for the disease to be openly discussed...impression was that they had clearer ideas about gene therapy in the following lessons. And let me tell you more...they had brilliant ideas on how to solve many therapeutic and pre-clinical problems! Who knows, perhaps...gamma-retrovirus or lentivirus-based ex-vivo gene therapy’ strategies to undergraduates, however you may ...

  3. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...lab from the University of Utah developed SapTrap. The SapTrap system uses golden gate assembly to produce...Ras oncogenes have been difficult to target therapeutically due in no small part to the central role Ras...restriction enzyme site and unique primer sites for more rapid screening by restriction digest and Sanger sequencing... sites, this technique also allows for easy and rapid targeting of complex elements to specific genomic...genome engineering method in bacteria that enables rapid, automated, and high-throughput genome editing is...(ssDNA) oligonucleotides (oligos), and thereby rapidly create desired allele combinations and combinatorial...et al. Metab Eng. 2012. PubMed PMID: 2232647   SapTrap, a toolkit for high-throughput CRISPR/Cas9 gene...

  4. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...organism. These features make AAV ideal for human therapies and in vivo laboratory experiments ranging from...in the nucleus and is a good option if you need rapid transgene expression, since you don’t have to wait...transduction independently of DNA synthesis." Gene therapy 8.16 (2001). PubMed PMID: 11509958. Vandenberghe...adeno-associated virus manufacturing." Human gene therapy 21.10 (2010): 1251-1257. PubMed PMID: 20649475....Efficient AAV Vector Production System: Towards Gene Therapy For Duchenne Muscular Dystrophy. InTech. doi: 10.5772...the titration of AAV-2 vector stocks."Molecular Therapy 6.2 (2002): 272. PubMed PMID: 12349826. Xiong, ...Various AAV Serotypes and Their Applications in Gene Therapy: An Overview.” Cells, 12(5), 785 (2023). PubMed...

  5. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ... islandicus CRISPR-Cas system. 5. Phage therapy Phage therapy uses phage to treat bacterial infections...promoters, light, or small molecules could achieve rapid, dynamic, and spatial control of CRISPR-Cas activity...bacterial pathogens possess CRISPR-Cas systems, phage therapy might not be as effective. This is where Acr proteins...enough that they could be engineered to “armor” therapeutic phages to protect them from pathogenic bacteria...activity is an important step towards CRISPR-based therapies and for more precise editing in the lab. References...Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development 9:390–401 . https...

  6. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...primary human cells. Now, scientists at Beam Therapeutics have found a way to improve editing in primary... and Head of Gene Editing Technology at Beam Therapeutics, used a previously developed bacterial selection...research! Base editing in primary human cells  Cell therapy approaches can use base editors to correct mutations...efficiently makes it a promising tool for T cell therapy development.  Reducing off-target editing in ABE8s...editing specificity, for example for potential therapeutic approaches, the authors recommend mRNA delivery...off-target deamination, an important attribute for therapeutic application. ABE8s demonstrate an overall improved...adenine base editors with increased activity and therapeutic application. Nature Biotechnology. https://doi.org...

  7. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ... agarose addition, we found it was possible to rapidly punch out cell clones with sterile truncated pipet...from a 10 cm dish in 15- 20 minutes- a far more rapid pace than using cloning rings. A picture of a 60...only warm but not hot- and still liquid- it is rapidly poured onto the middle of the plate, where it immediately...and partially dislodged from the agarose plug by rapid pipetting, as shown in the YouTube video. Before... appear to be yielding well-spaced clones. For rapidly growing cells we sometimes use TPP peelaway T-75...drop of serum to each well in the 48-well plate; rapidly growing cells such as HEK and CHO do not seem to...technique should be useful to scientists who wish to rapidly isolate individual cell clones with different expression...

  8. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...nanobodies that bind to coronavirus spike proteins (Wrapp et al., 2020), which sit on the surface of coronaviruses... read our secondary nanobodies blog post.  A rapid selection platform for generating sybodies against...developed a robust in-vitro protocol that enables rapid selection of synthetic nanobodies, called sybodies...for further characterization and analysis. In a rapid response to the COVID-19 pandemic, the Seeger lab...lab demonstrated they could use the protocol to rapidly identify and isolate sybodies targeting the SARS-CoV...BioRxiv.  https://doi.org/10.1101/2020.04.16.045419.  Wrapp D, De Vlieger D, Corbett KS, Torres GM, Wang N, ...single domain antibodies for the conformational trapping of membrane proteins. eLife 7: . https://doi.org...

  9. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org...dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https://doi.org/10.1038/s41434... Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089/hum.2009.169...Virus Serotype 9 (AAV9) Preparations. Human Gene Therapy Methods, 30(6), 214–225. https://doi.org/10.1089...Chemically and Functionally Distinct Vectors. Molecular Therapy - Methods and Clinical Development, 18(September...vector purification and formulation. Molecular Therapy, 12(1), 171–178. https://doi.org/10.1016/j.ymthe...

  10. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    Published
    Sept. 18, 2018, 1:33 p.m.
    ...Wanisch  Listen to the optogenetics podcast segment LOVTRAP is an optogenetic system that was recently developed...promoting domain. Each half was fused with one of the LOVTRAP domains. When both halves of the modified EB1 protein...system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing and Cas9 cleavage...popularity of single base-editors has resulted in the rapid expansion of the genome editing toolkit. However...technologies across several cell types is the lack of a rapid and quantitative editing assay. To help accelerate...has deposited a reporter plasmid that allows for rapid fluorescence-based quantification of base editors...

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

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...of the VGS process and use cases in Human Gene Therapy. Existing methods for AAV quality control Historically...Single-stranded Virus Next-generation Sequencing. Molecular Therapy - Nucleic Acids 4:e260 . https://doi.org/10.1038..., Paulk NK (2019) Fast-Seq: A Simple Method for Rapid and Inexpensive Validation of Packaged Single-Stranded...Viral Genomes in Academic Settings. Human Gene Therapy Methods 30:195–205 . https://doi.org/10.1089/hgtb...Moullier P, Ayuso E, Vandenberghe LH (2017) AAV-ID: A Rapid and Robust Assay for Batch-to-Batch Consistency ...Consistency Evaluation of AAV Preparations. Molecular Therapy 25:1375–1386 . https://doi.org/10.1016/j.ymthe.2017.04.001...

  12. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ... antibodies. This is particularly useful for therapeutic applications in vivo where endogenous anti-AAV...anti-AAV antibodies often neutralize the therapeutic efficacy of AAV vectors. How are exosome AAVs produced...interest special contexts, such as the delivery of therapeutic proteins to the brain which requires crossing...cells using exosome-associated AAV." Molecular Therapy 25.2 (2017): 379-391. PubMed PMID: 28082074. PubMed...vector as a novel gene delivery system." Molecular Therapy 20.5 (2012): 960-971. PubMed PMID: 22314290. PubMed...Overexpression of the Tetraspanin CD9." Molecular Therapy-Methods & Clinical Development 9 (2018): 278-287...

  13. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...retro-orbital) to intermediate (tail vein) Intraperitoneal Intraperitoneal (IP) injections are given in the peritoneal...drugs and viral vectors. Like IV injections, intraperitoneal injections require a little bit of practice...purposes, but mainly for the delivery of various therapies and viral vectors. This injection sounds very ... that require ingestion, such as experimental therapies or pain relief, straight to the stomach. This ...retro-orbital) to intermediate (tail vein) Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL ...

  14. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...used for many years for in vivo delivery and gene therapy and represent one of the most studied viruses. ...and very efficient introduction of DNA into therapeutically relevant, non-transformed mammalian cells. ...deficiency (most widely used so far) or for cancer gene therapy such as nonalcoholic steatohepatitis cancer. In...Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp 39–67 Ding Q, Strong A, Patel KM,... Zamyatnin AA Jr (2016) Viral vectors for gene therapy: Current state and clinical perspectives. Biochemistry... of Cas9-Specific Immune Responses. Human Gene Therapy 26:432–442 . https://doi.org/10.1089/hum.2015.087...

  15. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ...over two decades, attempts to answer it in five paragraphs, with examples. If that sounds ambitious, that...Science” described in Chapter 3, and of course the rapidly changing world of genome engineering via CRISPR...implications of using these technologies, but also the rapidity of the technologies’ adoption by the scientific... also spends some time on Addgene’s role in the rapid dissemination of these technologies, proposing that...are reading this review--in the current period of rapid flux, the chapter might still feel slightly dated...them wanting to learn more. There are individual paragraphs that may have warranted their own chapters, and...

  16. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...test for mouse iPSCs, tetraploid complementation, involves injecting tetraploid blastocysts with iPSCs...this approach is still not considered safe for therapeutic applications. piggyBac (PB) transposon: The PB...safer way to express reprogramming factors for therapeutic applications. Sendai viral vectors: Sendai virus...is considered a safer approach for developing therapeutic applications, but it can take a long time to ...induced pluripotent stem cells in research and therapy." Nature 481, no. 7381 (2012): 295-305. PubMed ... cell generation. Expert opinion on biological therapy, 10 2, 231-42. PubMed PMID: 20088717. PubMed Central...

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

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...CRISPR/Cas based COVID-19 testing methods.   Rapid detection of 2019 novel coronavirus SARS-CoV-2 using...incorporated into a lateral-flow readout slip that allows rapid detection.  Highlights: Cas13a is not catalytically...All-in-one dual CRISPR/Cas12a (AIOD-CRISPR) assay – for rapid, ultrasensitive and visual detection of novel coronavirus...operated, made accessible by the DIY-Bio movement. Rapid, field-deployable nucleobase detection and identification... D., Sharma, N., Kumar, M., … Maiti, S. (2020). Rapid, field-deployable nucleobase detection and identification..., & Alejandra, G. C. (2020). An ultrasensitive, rapid, and portable coronavirus SARS-CoV-2 sequence detection...

  18. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...diagnostic tools and therapeutics. In addition, the strategic use of whole phages as therapeutics, which is also...long-term library storage. After overnight growth, scrape up, pool and extract plasmid DNA directly from ...Nobrega, Franklin L., et al. "Revisiting phage therapy: new applications for old resources." Trends in...drug approval pathways for bacteriophage-based therapeutics."Frontiers in Microbiology 7 (2016). PubMed ...PMCID: PMC4728577. 4. Schmitz, Jonathan E., et al. "Rapid DNA library construction for functional genomic ...

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

    Type
    Blog Post
    Published
    July 7, 2017, 1:32 p.m.
    ... in therapy. The lungs are difficult, but other tissues like the retina have cell-based therapies underway...researcher. Darrell's ultimate goal - to develop therapies to treat disease in the lung. Chapter 2: Balancing...disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from the Kotton...

  20. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    Published
    June 30, 2015, 2 p.m.
    ...genome engineering - such as certain forms of gene therapy - avoiding the use of viruses is also important...of each construct. This new tool kit allows the rapid generation of stable transgenic cell lines which... CD19-directed chimeric antigen receptor (CAR) therapy. SB is a valuable tool and an alternative to CRISPRs..., et al. Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol...PubMed. 3. Singh H, et al. A new approach to gene therapy using Sleeping Beauty to genetically modify clinical-grade...
Showing: 41 - 60 of 419 results