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We narrowed to 223 results for: cas9 plasmid

Showing: 181 - 200 of 223 results

  1. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...every isolated plasmid sample and normalize each to fall within a 10 fold range. Plasmids that don't meet...A broader view of the whole plasmid To confirm the sequence of the plasmid, we examine three things: ...expected and do not affect plasmid function, we will update information on the plasmid’s page. Sometimes we’re...’re missing a piece of the plasmid There are regions of some plasmids that are particularly difficult ...some plasmids will not result in one complete, circular assembly. Most of the time, these plasmids are ...bottom of the plasmid page. Viral preps and pooled libraries In addition to regular plasmid sequencing,...sequence all plasmids in the pool but instead selectively sequence the variable portion of the plasmid in the...

  2. 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... 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...

  3. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi: https...

  4. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    ... 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 ...

  5. 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...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 ...thereby affect the titer of the virus. Additional plasmids will also affect experiment optimization. Before...

  6. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...that Addgene’s mission statement extends beyond plasmids, AAVs, and antibodies to our content, website,...mission is to enable that process. Our repository of plasmids, AAVs, antibodies, and data allows scientists ...changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the...Resources on the Addgene Blog Science careers Plasmids CRISPR Viral vectors Fluorescent proteins  ...

  7. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    Published
    May 4, 2021, 1:15 p.m.
    ...interest has been cloned into an expression plasmid, the plasmid can be introduced into host cells, such ...that the plasmids and sequences for antibodies are not shared. For instance, through open plasmid sharing...hundreds of useful variants of Cas9. By providing access to the plasmids encoding antibodies, scientists...Recombinant monoclonal antibodies are encoded in plasmids and produce a single antibody recognizing one ...synthetic genes that are typically expressed from a plasmid or from an integrated sequence in a stable cell...heavy chain genes of the antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an...laboratories and companies have already begun creating plasmids encoding affinity reagents. These include conventional...

  8. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...lead to issues such as recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots...library (How important is it that every single plasmid in your library is present, and how will you ensure...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...libraries often contain thousands, if not millions, of plasmids, they are generally electroporated into electrocompetent...

  9. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...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...

  10. 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...protein fusions Browse the Fluorescent Proteins Plasmid Kits ...

  11. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...Fluorescent protein plasmids Use FRET to study protein-protein interactions Deposit plasmids with Addgene! ...Tagged protein can easily be overexpressed from a plasmid but overexpression may interfere with the system...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even here...

  12. 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...

  13. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...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 ...

  14. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 ...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...

  15. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...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...recombination for amplification. The final purified plasmid stock should be analyzed by endonuclease restriction...-S). HEK293 cells are transfected with the AdV plasmid construct from step #1 and allowed to stay in culture...primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions ...

  16. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...the target locus, making them more flexible than Cas9/Cpf1. Some Cas13 enzymes prefer targets with a given...is mediated by ADAR2DD, not PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving...for viral delivery.  Find the RESCUE RNA editing plasmids here! Future directions Building on previous RNA...Science (2017):pii: eaaq0180. PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...550(7675) (2017):280-284. PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...27256883. PubMed Central PMCID: PMC5127784. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra...) (2016):270-273. PubMed PMID: 27669025. Find plasmids from this paper at Addgene. Gootenberg, Jonathan...

  17. 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...Experiment Deposit Your Plasmids with Addgene ...

  18. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...available (Weigel and Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...prefer Golden Gate assembly, there are a series of plasmids from the Dinneny lab that additionally allow for...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium... blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying plant genetic engineering...

  20. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ...versions of non-modular proteins like Cre. Could Cas9-DOG be developed soon to allow genome modification...23953120. PubMed Central PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan...2015): 1334-1341. PubMed PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer...
Showing: 181 - 200 of 223 results