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We narrowed to 230 results for: crispr c plasmids

Showing: 101 - 120 of 230 results

  1. Hot Plasmids: Spring 2025

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    Blog Post
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...Explore our empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing tagged proteins...and AAV plasmid sets launching later this year!  Find Addgene's own Lentiviral backbone plasmids here! ...integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger, L., Lim, J., Hiraizumi, ...Find the TurboCas plasmid here! Cenik, B. K., Aoi, Y., Iwanaszko, M., Howard, B. C., Morgan, M. A., Andersen... synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean, A., Chen, Y., Iyer...powerful tools in a scientist's toolkit is the empty plasmid backbone: the DNA vector that contains everything...

  2. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

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    Blog Post
    ...mixing and matching of parts. Gibson assembly meets CRISPR Gibson can be adapted to more complicated cloning...use of Gibson assembly combined with the popular CRISPR technology and is described in a recent publication...second example of using Gibson Assembly along with CRISPR was also described recently (Jiang et al. 2015)...chromosome (up to 100 kb) were specifically cut out via CRISPR and then assembled into a vector using Gibson assembly...homology with the bacterial chromosome fragments. CRISPR cutting was used to circumvent the need to PCR ...Ligation Independent Cloning (SLIC) in a previous Plasmids 101 post. Although SLIC may be more cost effective...Wang A, Li K, Wang B, Jin S, Reiser M, Lockey RF. CRISPR/Cas9 nuclease cleavage combined with Gibson assembly...

  3. 22 Hot Plasmid Technologies from 2014

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    Blog Post
    ...information on these new CRISPR screening tools, see our CRISPR/Cas Plasmids: Pooled Libraries webpage...CRISPR-Cas resources at www.addgene.org/CRISPR/ to browse CRISPR plasmids, watch informational videos, download...Tet-inducible CRISPR activators and repressor plasmids. (Kearns et al., Development. 2014..) CRISPRs for Xenopus.... 2014..) Our CRISPR-Cas collection of plasmids updates frequently, so visit our CRISPR pages often to... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...vectors and sgRNA libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism...Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening. Also, check out our updated CRISPR-Cas ...

  4. Pooled Library Amplifications

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    Blog Post
    ...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...Amplification Type Link to Protocol CRISPR Knockout Human CRISPR Knockout Pooled Library (Brunello) ...Amplification Protocol (PDF, 88.2 KB) CRISPR Knockout Human CRISPR Knockout Pooled Library (GeCKO v2) ...Protocol (DOCX, 17 KB)   For CRISPR libraries, Genome-scale CRISPR-Cas9 knockout and transcriptional... Protocol (DOCX, 15 KB) CRISPR Activation Human Genome-wide CRISPRa-v2 Libraries 104,540–209,080...Re-Amplification Protocol (PDF, 71 KB) CRISPR Activation Human Subpooled CRISPRa-v2 Libraries 12,285–20,145 ...

  5. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...the ability to distribute plasmids with the piggyBac™ transposon. These plasmids, when combined with a source...donor plasmid carrying the DNA edit of interest, along with a site-specific nuclease (such as CRISPR/Cas9...piggyBac™ Transposon Catch up on Everything CRISPR with Our CRISPR Guide Deposit Your piggyBac™ Vectors! ...useful resource. While Addgene cannot distribute plasmids with the piggyBac™ transposase itself, please ...genome fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur... to the development of site-specific nucleases (CRISPR/Cas9, TALENs, and ZFNs). However, several challenges...piggyBac™ transposon system can be combined with CRISPR/Cas9 to efficiently perform genetic manipulations...

  6. Hot Plasmids - February 2022

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    Blog Post
    ...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'... Penn State Protein Ladder Engineered miniature CRISPR-Cas system for mammalian genome regulation and ...gene perturbation by: Gabrielle Clouse While many CRISPR-based genetic screening methods have primarily ...Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system is a cost-effective alternative... ladders. Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60...

  7. Technique: Probe Phage Genomes for Host Binding Proteins

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    Blog Post
    ...alternating between 5 min at -20°C (or -80°C) followed by 5 min at 37°C (each repeated 5 times) works well...pool and extract plasmid DNA directly from the colonies. Transform the resultant plasmid DNA into E. coli...DNA bases.  Ligate and transform your resultant plasmid libraries into chemically competent E. coli and... you’re using a pET vector) and antibiotic for plasmid selection. Remove air bubbles between the membrane...they expressed well, the proteins encoded by the plasmids in the library will remain attached to the membrane...PMC2258639. Resources on the Addgene Blog Learn How CRISPR Can Be Combined with Phage for Antimicrobial Purposes...Systems to Probe Protein-Protein Interactions Use CRISPR For Genome-Wide Screens Resources on Addgene.org...

  8. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

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    Blog Post
    ...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Human whole genome...Susanna Bachle Figure 1: Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu...'s lab at Peking University has developed a new CRISPR screening method that makes use of guide RNAs with...false-positive rate discovery rate, conventional CRISPR screens require a low multiplicity of infection...attached fluorescent protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry...

  9. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

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    Blog Post
    ...Addgene Blog First time CRISPR user? Check out our tips for 1st time CRISPR users Learn how to Choose...Here Brush Up on Your CRISPR Background with Addgene’s CRISPR Guide Page ...resolution, the Xu lab created a dCas9-based CAPTURE (CRISPR Affinity Purification in situ of Regulatory Elements...at hundreds of loci at once (Liu et al., 2020). CRISPR Affinity Purification in situ of Regulatory Elements...CAPTURE 2.0 to be used with primary cells. Find plasmids for CAPTURE 2.0! When compared to the original... crispr...conformation capture (3C) based methods, such as 5C or Hi-C (Han et al., 2018). Think of 3C methods as taking ...

  10. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...View Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments Get Ready-to-Use CRISPR Lentivirus...steps in the CRISPR experimental process is validating edits. Regardless of which CRISPR genome editing...discussed in CRISPR 101: Validating Your Genome Edit, there are a variety of options for CRISPR genotyping...Perform Genome-Wide Screens with CRISPR Generate Mouse Models with CRISPR/Cas9 Additional Resources on .... Bell, Charles C., et al. "A high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations...target gene. The validation process, also known as CRISPR genotyping, is critical to demonstrating causal...biology. It is key to address these concerns as CRISPR use grows across the life sciences and to establish...

  11. Generating Mouse Models Using CRISPR/Cas9

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    Blog Post
    ...Addgene.org Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse ...Crossing Mice. My first CRISPR mouse experiment Everything changed with the advent of CRISPR in 2013 (Cong et...oligonucleotide or plasmid as the repair material if needed (for knock in models). CRISPR mouse model basics...using CRISPR for mouse genome engineering From the very start, generating mouse models using CRISPR is easier...Recent Post for Tips on CRISPR Gene Editing in Mice Disadvantages of using CRISPR for generating mouse models...remains to be seen whether CRISPR is as robust as conventional gene targeting. CRISPR tools were recently used... by guest bloggers, Wenning Qin and Haoyi Wang. CRISPR/Cas9 is revolutionizing the mouse gene-targeting...

  12. Developing a cold tolerant E. coli using specialized chaperones

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    Blog Post
    ...defined between a fairly nippy 20°C to a decidedly uncomfortable 45°C). Their more adventurous cousins ...of growth, say around 37°C, is a far cry from their native conditions of ~0°C, which drastically affects...transformed mesophilic, for example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes...granules. We overcame this problem by engineering a plasmid which contained cold tolerant chaperones (the Cpn...pMBP-LbCas12a and pC013-Twinstrep-SUMO-huLwCas13a plasmids from Addgene, which would potentially allow a ...temperature conditions, thereby enabling efficient CRISPR-Cas mediated editing for psychrophilic systems ...biology blog posts Resources on Addgene.org Find plasmids for your synthetic biology experiment Read our...

  13. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ...move antibiotic resistance genes between plasmids or from plasmids to the bacteria’s genome. In humans, if...encodes a Type I-F CRISPR-Cas system. This system has four major components: 1) a CRISPR RNA (crRNA), 2) ... its preferred target integration site: TNA(C/G)TTATAA(G/C)TNA. Tol2 can deliver 10-11 kB to mammalian...activity (Voigt et al., 2016). Sleeping Beauty plasmids  piggyBac Although its name suggests otherwise...codon-optimized wild-type piggyBac transposase.  piggyBac plasmids    Tol2  Tol2 was the first reported active DNA...increase in transposition activity  Tol2 transposon plasmids    Applications of transposons Now that you’ve...sets the stage for combining the super powers of CRISPR and transposons to allow for the targeted delivery...

  14. Plasmids 101: Modular Cloning

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    Blog Post
    ...assemble and test many variables all at once. CRISPR CRISPR arrays and libraries depend on many gRNAs delivered...As a plasmid repository, we are no strangers to cloning. If you’re reading this, then chances are you...etc. MoClo is especially useful for more complex CRISPR experiments (e.g., many guides/parts delivered)...delivered) in certain model organisms. In mammals, most CRISPR libraries are assembled and delivered virally, ...NEB’s DAD Interview with Nicola Patron on MoClo Plasmids 101: Golden Gate Cloning ...References and Resources References Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. A modular cloning...

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

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    Blog Post
    ...doi.org/10.1038/mt.2009.280 Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski...increasing the size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split...sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains: 400bp homology arms to ...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....2010.205 Trapani I, Colella P, Sommella A, Iodice C, Cesi G, Simone S, Marrocco E, Rossi S, Giunti M, ...

  16. Twenty Years of Sharing: Addgene's Viral Vector Service

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    Blog Post
    ...realized we could apply the lessons learned from our plasmid lab to develop a reliable and robust high quality...offer many resources for producing vectors from plasmids — including the protocols we use in our lab — ...while viral vectors are not as easy to ship as plasmids, they still have a global reach. From 2016–2023...vectors in our repository are produced from only 413 plasmids containing a genetic payload. While AAV9 and AAV1... may recall our first 20th anniversary story on CRISPR, where we dove fairly deeply into the different...having reliable ready-to-use vectors made from plasmids in the Addgene catalog would be broadly useful...conditions. Sharing our experiences Addgene’s plasmid repository gave us the experience to launch a high-quality...

  17. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources....   C. roseus is not a model organism such as Arabidopsis or tobacco. Studying gene function in C. roseus...its function. In nature Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...in tissue culture. This method is commonly used in C. roseus, but processes specific to photosynthetic ...Unver and Budak, 2009). This method works well in C. roseus. A common method for overexpressing genes ...

  18. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    Blog Post
    ...with some plasmids. You just need three plasmids to start making AAV:  the packaging plasmid which contains... transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids have two ...of an AAV transfer plasmid. Parts shown in green are key elements of an AAV plasmid. Parts shown in grey... elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal ...reporter like GFP, some smaller CRISPR-Cas proteins such as SaCas9, CRISPR gRNAs, or your gene of interest...transfer plasmids have a promoter, but this element isn’t required when the transfer plasmid harbors non-coding...Transfer Plasmid video!   Now you’ve got a basic understanding of some common AAV plasmid elements!...

  19. Lentiviral Vector Uses and Overview

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    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Addgene Blog Your Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale...Lentiviral vectors are the vector of choice for many CRISPR applications, and they’ve also had success in clinical...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 ...

  20. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides...AAV plasmid standard and AAV transfer plasmids. It’s easy to contaminate a workspace with plasmid, not...bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination...going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs which are...AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate...mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple ways to ...
Showing: 101 - 120 of 230 results