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We narrowed to 215 results for: crispr cas9 plasmids

Showing: 181 - 200 of 215 results

  1. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...knowledge changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in...Resources on the Addgene Blog Science careers Plasmids CRISPR Viral vectors Fluorescent proteins  ...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 ...

  2. 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...

  3. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...a 1st Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian...48-well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western ...; more recently we have also been using Addgene CRISPR vectors to generate cell lines with knockouts of...Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...

  4. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...RNAi to knock-down expression of your target or CRISPR/Cas9 to create mutations that essentially eliminate...gene for your protein has been mutated using CRISPR/Cas9. You check the mutation and it causes a large...interest endogenously and transfect them with a plasmid encoding your protein of interest. Then you should...target protein in a Western blot. You acquire a plasmid encoding your protein of interest, known as Gene... In parallel, you transfect cells with another plasmid not encoding this protein, which we will call Negative...band from the cells transfected with the other plasmid. Your antibody works! Multiple (Independent) Antibodies...Antibodies blog post!  Resources on Addgene.org Find plasmids encoding antibodies at Addgene Stay tuned for ...

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

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has revolutionized ...A, Bassik MC (2016) Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes. Nat Biotechnol...our CRISPR blog posts Resources on Addgene.org Find tools for mammalian RNAi Browse all plasmids for...unprecedented ways. With the meteoric rise and diversity of CRISPR technologies, one might incorrectly relegate RNAi...straightforwardness of RNAi is unmatched—unlike CRISPR experiments that require exogenous protein, successful...only requires the presence of dsRNA. In plants, CRISPR technologies are predominantly used to disrupt ...function at the level of DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic...

  6. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...min. Using these lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here. Listen to the Hot Plasmids Podcast!  MarathonRT...’s lab at Yale University recently deposited a plasmid for expressing a novel reverse transcriptase (RT...polymerase processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA. 2018 PubMed PMID:29109157... cell surface. Their toolkit includes a set of plasmids each containing an anti-GFP nanobody fused with...

  7. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ... of CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus.... The continued development of more and smaller CRISPR tools is an exciting area for future AAV application...

  8. Meet the New Addgene Board Members!

    Type
    Blog Post
    Published
    April 28, 2020, 1:14 p.m.
    ...Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function ...over 100 requests for the plasmids and the lab eventually deposited these plasmids at Addgene. While deposits...The mission to share critical reagents such as plasmids and viral stocks for researchers around the world...

  9. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid maps far easier...easier for you to share plasmid information with colleagues, or explain plasmid features to a lab member... on the main plasmid page under the "Resource Information" section. Analyze your plasmid sequence For ...over from our users: "Please, please improve your plasmid maps!" After thoughtful design, vetting, and tweaking...tweaking, we’re excited to announce that our plasmid and sequence displays are now powered by GSL Biotech's...software and extensive feature library, our updated plasmid and sequence displays are now much easier to interpret...how they’ll make it easier for you to find the plasmids you need. We'll be monitoring and improving the...

  10. Luminescent Imaging with Nano-lanterns

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

  11. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...Read all CRISPR blog posts Resources on Addgene.org Browse our CRISPR interference plasmids Find bacterial...bacterial CRISPR plasmids Read our CRISPR guide ...Mobile-CRISPRi into bacteria. Top: A Tn7 transposon containing the CRISPRi components and a plasmid with...ready to move Mobile-CRISPRi into your bacteria of interest? Find Mobile-CRISPRi plasmids at Addgene! References...about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko, CRISPRi, and CRISPRa...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria..., tnsC, tnsD, and tnsE), and a plasmid that contains the Mobile CRISPRi cassette flanked by left and right...

  12. 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...high titer virus for transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest...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 ...

  13. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even...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...

  14. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...Photosactivatable Fluorescent Proteins Use Light to Control CRISPR Genome Editing Choose the Brightest Fluorescent...Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids from the Tsien Lab Browse the Michael Davidson...

  15. 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...sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing access to the ...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...

  16. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources page...-1 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To find... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting... Find the plasmids at Addgene! CRISPR RNP electroporation and AAV donor infection (CRISPR-READI) enables...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...

  17. 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.
    ...sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to the...increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split...single step and serve as donors for two consecutive CRISPR-mediated HDR events which results in the integration....org/10.1172/jci34316 Bak RO, Porteus MH (2017) CRISPR-Mediated Integration of Large Gene Cassettes Using...therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out the Addgene...

  18. Adapting Toehold Switches to Detect Zika Virus

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

  19. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ... primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions...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...

  20. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources ...acssynbio.0c00192   The CRISPR corner By Jennifer Tsang New CRISPR plasmids are always being added to...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...article: CRISPR-based editing in organoid Biosensors for detecting strigolactones The CRISPR Corner New...system, showing the workflow for inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy...New ready-to-use viral preps New toolkit for CRISPR-based genome editing in organoids By Ashley Waldron...
Showing: 181 - 200 of 215 results