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We narrowed to 170 results for: cas9 expression plasmid

Showing: 141 - 160 of 170 results

  1. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. .... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing efficiencies...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities...

  2. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of ...capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response... capacity (up to 36 kb!), long-term transgene expression and negligible toxicity. However, their production...down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular...adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene of interest...how to generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide...2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain...

  3. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...extra level of safety, they also require a fourth plasmid. The requirement for additional DNA can impact ...

  4. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ... Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected...Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org...

  5. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...Nathaniel Roquet from the Lu Lab at MIT developed plasmid based systems that report the order in which specific... action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “... plasmid... system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable...

  6. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the key tools now providing access...plans and in particular, that they all share the plasmids created under this programs via Addgene. New organisms...snail can be manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated...sexual reproductive life cycles. One can inject plasmid DNA in the eggs to get transgenic animals. They...-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod germline by injection into ...

  7. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ...has a low quantum yield, and when transiently expressed in cells, generates much less signal than comparable... endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging ...: 1262. PMID: 23232392 PMCID: PMC3535334 Find plasmids from this paper at Addgene. Takai, Akira, et ...4352-6. PMID: 25831507. PMCID: PMC4394297 Find plasmids from this paper at Addgene. Suzuki, Kazushi, ... 13718. PMID: 27966527 PMCID: PMC5171807 Find plasmids from this paper at Addgene. Additional Resources...All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein ...

  8. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ...interest (or the whole genome), they knock down gene expression temporarily. RNAi libraries are especially useful... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi: https...

  9. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...Browse fluorescent protein plasmids and resources See all Jaffrey lab plasmids ... cellular processes by transforming cells with plasmids encoding aptamer-tagged RNA and later introducing...aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex. Broccoli...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

  10. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...generation of multiple edits or implementation of expression systems to accurately mimic a disease phenotype...CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the genome Tips for a first...resources on Addgene.org CRISPR Guide CRISPR Plasmids and Resources CRISPR History and Development ...

  11. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...Turquoise (CeNL; Plasmid #85199), Yellow (YeNL; Plasmid #85201), Orange (OeNL; plasmid #85202) and Red ... Researchers have been sharing plasmids ever since there were plasmids to share. Back when I was in the...specific strategies and tactics for getting the plasmids he wanted. Addgene makes it so much easier to ...endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting for endogenous...kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk...reporter was used to sort successfully infected/expressing cells by flow cytometry. The isolated cells were...CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into mouse livers...

  12. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...Drawbacks: Transgene expression varies with this approach, ranging from ~10% expression in primary human ...this AAV successfully mediated transduction and expression of the full-length transgene in vitro and in ...genomes complement each other to restore the full-expression cassette. AAV’s natural ability to reassemble...overlapping split vector has equal or higher transgene expression when compared to using a single over-sized vector...could have a big impact on the rate of transgene expression with an overlapping AAV since both of these variables...sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview: The hybrid...

  13. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...Unpublished or pre-publication plasmids": Unpublished plasmids or plasmids that will not be published are...and entering a plasmid name,plasmid type, and plasmid description/experimental purpose (right). On the...with your plasmids, and adding all the plasmids at once increases the likelihood that the plasmids are given...easy for you to share plasmids. To achieve this goal, we will archive any plasmids you've deposited with...Spreadsheet". Start your plasmid deposit There are two options to begin your plasmid deposit. Figure 2: Choose... Choose to submit plasmid(s) online or by using a spreadsheet. The plasmid information can be entered ...than 10 plasmids but we recommend using the spreadsheet option for deposits of 10 or more plasmids.   ...

  14. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ... Check out All the Plasmids from the Collin's Lab Find Synthetic Biology Plasmids for Your Next Experiment...amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic reader...Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from the publication at Addgene 2. Green, Alexander...switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939. PubMed PMID: 25417166...Experiment Deposit Your Plasmids with Addgene ...

  15. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus...for model organisms.    Q11: How consistent is expression is when injecting CAP-B10 or CAP-B22 in marmosets...

  16. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available, or...of interest when expressed at endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly...flow cytometry, and mRNA expression.   If your protein of interest is expressed at similar levels in ... involves the transient transfection of expression plasmids in a given cell line. For flow cytometry, ...useful to produce a sample with mixed expression, with the expression measurable through an epitope tag such...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...caused by overexpression. Additionally, if the cell line used has endogenous levels of expression, this can...

  17. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...activity. To drive selective A → T editing, they co-expressed Polη, a translesion synthesis polymerase known...inserted the deaminase into the C-terminal domain of Cas9, which narrowed the editing window and reduced A...site. Like in most other base editor designs, the Cas9 nicks the non-edited strand to encourage the cell...reporters Resources on Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our.... gGBE consists only of engineered MPG fused to nCas9. First, MPG removes the G base from DNA to create...

  18. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...nuclease-deficient Cas9 can bind loci in the genome that are defined by a gRNA. Since Cas9 is a protein, a... "Live cell imaging of phosphoinositides with expressed inositide binding protein domains." Methods 46.3...protein fusions Browse the Fluorescent Proteins Plasmid Kits ...

  19. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...for mammalian RNAi Browse all plasmids for RNAi Check out Addgene's plasmids and resources for plant biology...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence...harvested instead of microscopic isolation. The plasmids required to conduct an APT-based RNAi experiment... target sequence, as well as essential control plasmids. Many thanks to our guest blogger Robert Orr...Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...

  20. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ...popular split-FP plasmids from our catalog to help you find the right split-FP plasmid for your experiment... vitro, and can be used for measuring protein expression or localization in cells, or even single-particle...below, and click the Article link to find all the plasmids from the associated paper:   Color Base FP...and experimental design. We hope you find these plasmids useful. Good luck, and happy experimenting! References...Split-horseradish peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015...
Showing: 141 - 160 of 170 results