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  1. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ... which. Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a...tip! In CRISPR, the non-target strand is the DNA strand that does not base pair with the gRNA. Base editors...blog CRISPR 101: Cytosine and Adenine Base Editors Read about four base editor reporters CRISPR 101: ...Editing CRISPR 101: Cas9 Structural Biology Additional resources on Addgene.org Browse CRISPR plasmids...editing Get a broad overview with our CRISPR Guide Check out our CRISPR 101 ebook ...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...base editing tools First, let’s briefly review how CRISPR-derived base editing works in general. Base editors...
  2. CRISPR Plasmids - Prime Edit

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    ...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Prime... CRISPR Prime Edit CRISPR Plasmids: Prime Edit Browse CRISPR Plasmids By Function ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...find a gRNA vector based on expression system, promoter, the type of gRNA (e.g., pegRNA, epegRNA, nicking...Publication Empty Prime Editing gRNA Vectors A selection of empty gRNA vectors suitable for prime editing...using a prime editing guide RNA (pegRNA). As with a typical gRNA, the pegRNA is designed with a spacer that... . Browse, sort, or search the tables below for CRISPR prime editing plasmids. Plasmids are available ...
  3. Hot Plasmids and Viral Preps - March 2021

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    Blog Post
    ...very fast CRISPR, is a light-inducible CRISPR system developed by Taekjip Ha’s lab where Cas9-gRNA complex...       The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To...To find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources...auxin inducible degron system for C. elegans New CRISPR plasmids New items from the viral service Epigenetic...Saptrap cloning systems to generate AID* tagged CRISPR knock-in repair constructs. These repair constructs...tagged AID* into genomic sites of interests via CRISPR techniques. This tool-kit includes an array of ...Cas9 cleavage can induce DSBs within minutes. New CRISPR plasmids for Borrelia burgdorferi from Christine...
  4. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

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    Blog Post
    ...Most popular methods for CRISPR activation CRISPR activation uses dCas9, a CRISPR protein variant lacking... blog Browse our many CRISPR blog posts Get the basics of CRISPR in the CRISPR 101 blog series Resources...making it a common choice for CRISPR activation. Newer methods for CRISPR activation While SAM, SunTag,...compared to other CRISPR activators, but their initial results show promise for CRISPR-based gene activation...Resources on Addene.org Read Addgene's CRISPR Guide Find CRISPR plasmids for your research ...Prior to the discovery of CRISPR/Cas systems, gene activation across multiple loci was an arduous process...activation seem next to impossible. The development of CRISPR/Cas systems, however, greatly improved the simplicity...
  5. CRISPR 101: Cas9 vs. The Other Cas(s)

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    Blog Post
    ...resources on the Addgene Blog CRISPR 101: Which Cas9 do I choose for my CRISPR experiment? The PAM requirement...expanding CRISPR beyond spCas9 Cas proteins More resources on the Addgene Website CRISPR Plasmids ...engineering workhorse You rarely hear the word ‘CRISPR’ without shortly thereafter hearing ‘Cas9’. The...good reason. It is guided by an RNA molecule, the gRNA, to a complementary site in the genome where it ...targeting. RNA targeting Cas enzymes Move over Cas9, CRISPR isn’t just for DNA anymore! Cas enzymes have been...Deletions in Human Embryonic Stem Cells Using Type I CRISPR-Cas. Molecular Cell, 74(5): 936-950.e5 (2019). ...is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...
  6. Hot Plasmids and Viral Preps - January 2021

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    Blog Post
    ...yeast CRISPRa plasmids here Reduce the scale of your CRISPR screens by using this lentiviral CRISPR human...for use in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items ...Hypercompact genome editor CRISPR-Casɸ By Melina Fan The Doudna Lab adds to the CRISPR-Cas toolbox with their...sciadv.abb7438 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To...To find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources...chemically inducible CRISPRa system in yeast is mediated by recruitment of MS2-functionalized gRNAs.  Find the...their discovery of CRISPR-Casɸ, a hypercompact genome editor from the Biggiephage clade of huge phages. ...
  7. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

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    Blog Post
    ...targeting range of CRISPRCas9 by engineering SpCas9 Article contributed by Shreya Vedantam CRISPR/Cas9 is widely...2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method to tag endogenous proteins in...and other tags (Yesbolatova et al., 2019) using CRISPR. This toolkit makes it easy to tag your protein...call the LADL Anchor. Using specifically designed gRNAs, they can then recruit these LADL Anchors to genomic...
  8. FlyCRISPR

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    ...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Harrison Lab - Wildonger... CRISPR flyCRISPR Lab CRISPR Plasmids flyCRISPR Plasmids Available from Addgene You may also...for generating gRNAs 106.5 KB flyCRISPR website contains additional information CRISPR Optimal Target ...Browse CRISPR Cas9 gRNA plasmids for drosophila research from the O'Connor-Giles, Wildonger, and Harrison...Target Finder Identifies gRNA target sequences from an input sequence and checks for off-target binding. Currently...described in: Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Gratz SJ, Cummings AM...Wildonger Lab - O'Connor-Giles Lab email: flycrispr@gmail.com We have found that a variety of Cas9-mediated...
  9. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...up-to-date information. Check our CRISPR guide and blog to learn more about CRISPR and the most recent developments...prep from the Addgene viral service. CRISPR technology lentiCRISPR v2 (Plasmid #52961) This is a 3rd generation...original lentiCRISPR v1. Special note from the Zhang lab: We are constantly improving our CRISPR reagents...by delivering shRNA or miRNA. The ever expanding CRISPR field makes wide use of retroviral delivery systems...systems, especially lentiviral vector-based CRISPR pooled libraries that allow functional screening of ...expression, gene knockout, and selection marker removal. CRISPR technology pdCas9-humanized (Plasmid #44246) Expression...Vector provided by the Qi lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral...
  10. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...Limitation for CRISPR Delivery Genome-wide Screening Using CRISPR Performing In Vivo CRISPR Screens Using...development and improvement of CRISPR-based technologies over the past decade. CRISPR screens are easily customizable...resources at addgene.org CRISPR pooled libraries Guide to pooled libraries CRISPR guide AAV guide ... interference, activation, and knockout. Most CRISPR screens rely on lentiviral systems, a fact supported...supported by the wide array of lentiviral-based CRISPR libraries available at Addgene. The large packaging... AAVantageous!       In vivo, in vitro, ex vivo CRISPR screens can be carried out using a few different... process of these screens is the same: Deliver CRISPR library to cells of interest. Select for cells ...
  11. CRISPR History and Development for Genome Engineering

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    ...like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR revolution shows no...information on CRISPR, check out Addgene’s CRISPR Guide and CRISPR blog posts , or find CRISPR plasmids for... CRISPR CRISPR History CRISPR History and Development for Genome Engineering You may also like...sequencing, CRISPR is a robust system for genome-wide screening. CRISPR Technological Advances CRISPR technology...with paired gRNAs to lower off-target cleavage frequency. Truncated gRNAs: Truncated gRNAs display less...finger nucleases (ZFNs) and TALENs. CRISPR: An Adaptive Immune System CRISPR (Clustered Regularly Interspaced...prokaryotes used CRISPR as part of an adaptive immune system - utilizing various CRISPR-associated (Cas...
  12. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...techniques such as CRISPR are easier to use and are more efficient at inactivating genes. CRISPR can target a...have developed new CRISPR-based methods for various knock-ins, discussed in our CRISPR 101 eBook. It’s ...genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems, these systems ...systems remain valuable, especially in cases where CRISPR cannot be used (e.g. there are no suitable PAM sequences...interest is difficult to target specifically with a gRNA). Be sure to keep these techniques in mind when ... to create conditional floxed alleles. Although CRISPR is very good at making knockouts, knocking in large...thirty years ago. As traditional knockout and new CRISPR tools continue to be perfected, the production ...
  13. Plant Plasmids and Resources

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    ... Addgene! CRISPR Please refer to Addgene's CRISPR Guide for a general introduction to CRISPR technology... The MoClo CRISPR/Cas Toolkit for Plants includes CRISPR/Cas nucleases, base editors, gRNA backbones, ...like... Plant CRISPR resources Fluorescent Protein Guide Addgene Blog: Plant Biology CRISPR Cloning Transcription...variants of Cas9 and empty gRNA backbones. A few examples of useful plant CRISPR resources include: A Multipurpose...assembly of one or more gRNA expression cassettes - Qi-Jun Chen. Cas9 and gRNA backbone for use in rice...technology or the Plant CRISPR Resources for a full selection of plasmids expressing different functional variants...truncatula , wheat, and barley - Daniel Voytas. A CRISPR/Cas9 Toolkit for use in a variety of plant species...
  14. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ... out our CRISPR Featured Topic Page Learn about RNA Targeting with Cas13a Learn about CRISPR DNA Base ...Resources on Addgene.org Browse All CRISPR Plasmids Check out our CRISPR Guide Pages ...enzymes are quickly becoming major players in the CRISPR field. Just a year after Feng Zhang’s lab identified...C2c2) (Abudayyeh et al., 2016) as a RNA-targeting CRISPR enzyme, they adapted Cas13b for precise RNA editing...programmable A to I (G) replacement) is the first CRISPR tool for RNA editing. Two years after that, the...edit to be made when multiple As are present in the gRNA spacer, as ADAR will preferentially edit an adenine... targeting work with Cas13, REPAIR is the first CRISPR-based system to enable precise RNA editing. The...
  15. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ...editing Read more about CRISPR Learn all the basics about CRISPR in our CRISPR 101 blog series Resources...Through molecular biology teamwork. The guide RNA (gRNA) specifies the editing target site on the DNA, the... Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and Beyond meeting earlier this month...Addgene.org Find base editors for your research Read our CRISPR guide Find plasmids for your research ...
  16. dTAG - You're it!

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    Blog Post
    ...Addgene.org Browse all CRISPR plasmids CRISPR plasmids for protein tagging Find validated gRNAs ...Addgene Blog Easi-CRISPR: Generating Knock-In and Conditional Mouse Models CRISPR 101: Cas9 Nickase Design... selective degradation of FKBP12F36V-BRD4 using CRISPR/Cas9-mediated knock-in, and to evaluate in vivo...ENL-FKBP12F36V and endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and...-MELK and endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, ...were engineered to express YY1-FKBP12F36V using CRISPR/Cas9-mediated knock-in. Featured molecule: dTAG...
  17. Qi Lab CRISPR Page

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    ...Addgene You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts We have recently... CRISPR Qi Lab CRISPR Plasmids Qi Labs CRISPR Plasmids...CRISPR CRISPRi Cas9 gRNA plasmids from the Qi lab CRISPR...or human cells ( Qi et al. ). This CRISPR interfering system (CRISPRi), works independently of host cellular...uniqueness of CRISPRi, as compared to several recently published works on applying the wild-type CRISPR system... fusion expression plasmid 44248 pgRNA-humanized A customizable gRNA expression plasmid for use with either...use editing bacterial genomes 44251 pgRNA-bacteria A customizable gRNA expression plasmid for use with the...
  18. Google Forums Round Up: First Impressions of NgAgo

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    Blog Post
    ... Request NgAgo Find CRISPR Resources Catch Up on Your CRISPR Background ...genome engineer sharing the stage with much-lauded CRISPR-Cas9 is DNA-guided endonuclease NgAgo! We'll discuss...development before it can truly be pitted against CRISPR head-to-head.  Exciting features of NgAgo First...which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and NgAgo genome engineering methods have ...Yourself On the other hand, the widespread success of CRISPR-Cas9 across multiple model organisms (3) lies in...transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA- and Cas9-expression...had tested and compared NgAgo head-to-head with CRISPR-Cas9. The survey had two sections, the first section...
  19. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...blog Browse our CRISPR blog posts Download our CRISPR cheat sheet Download our CRISPR eBook Resources...out our comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...recent papers describe the development of CRISPR activation (CRISPRa) tools for neuroscience, including transgenic...neuroscience research. What is CRISPR-based activation? To summarize, CRISPRa uses a nuclease-dead variant...). Created with BioRender.com. Before using gRNAs in vivo, they should be screened for activity in...molecular gymnastics for sensitive applications such as CRISPR screening. The plasmids used by the Qi Lab for ....     References Liu, Yanxia, et al. "CRISPR activation screens systematically identify factors that drive...
  20. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...Addgene.org Catch up on your CRISPR background with our CRISPR Guide Browse CRISPR plasmids for base editing...Early CRISPR applications were often limited by the low editing efficiency of homology-directed repair...nickase) fused to a deaminase that makes the edit A gRNA targeting Cas to a specific locus A target base ... indels were observed than point modifications. CRISPR base editing persists through multiple cell divisions...editing based on Cas9 off-target activity.  Find CRISPR base editing plasmids at Addgene Improving cytosine...which you can learn more about in this blog post: CRISPR 101: RNA Editing with Cas13. Finally, the development...adenine deaminase TadA-8e for efficient and specific CRISPR-based cytosine base editing. Nature Biotechnology...
  21. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    ...screening, and validation of CRISPR libraries. The Genome Engineering using CRISPR/Cas Systems Forum- Moderated...Lentiviral CRISPR Libraries- A nice overview of what to consider when using pooled lentiviral CRISPR libraries...Addgene, including cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries at Addgene...but we do distribute several of our most popular CRISPR pooled libraries and one of our most popular barcoding...Resources and references References Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening...Practical Considerations for Using Pooled Lentiviral CRISPR Libraries. Curr Protoc Mol Biol. 2016 Jul 1;115..., Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening...
  22. Validated gRNA Sequences

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    ...like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA sequences... Genome Engineering CRISPR Validated gRNA Sequence Datatable Validated gRNA Target...PAM sequence). Which CRISPR application is this gRNA sequence compatible with? CRISPR knockout experiments...Does a given gRNA sequence exactly match your genomic target? Variation between a given gRNA sequence and...pyogenes, S. aureus etc.) was this gRNA sequence designed for? A given gRNA sequence may only be compatible...Addgene's CRISPR Plasmids and Resources page have been used to indicate the Cas9 application the gRNA was designed...details and outcomes. Submit Validated gRNA Sequences Validated gRNA sequences can be added to this table...
  23. Zhang Lab's CRISPR Frequently Asked Questions

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    ... Genome Engineering CRISPR Zhang Lab CRISPR Plasmids Zhang Lab CRISPR FAQs Zhang...Zhang Lab's CRISPR Frequently Asked Questions You may also like... Zhang Lab CRISPR plasmids General CRISPR...using the lab's CRISPR technology. Visit the Zhang Lab page and forum to learn more. CRISPR Design FAQs Homologous...the Bench FAQs CRISPR Design FAQs Should I use wildtype or double nickase for my CRISPR genome engineering...Dr. Le Cong, the first author of a seminal CRISPR article from Dr. Feng Zhang's Lab, has assembled a ... lab's CRISPR technology. Genome... CRISPR Plasmids Dr. Le Cong, the first author of one of the seminal articles from Dr. Feng Zhang's Lab...
  24. Jaenisch Lab CRISPR Plasmids

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    ...Lab CRISPR Plasmids Available from Addgene You may also like... CRISPR Guide CRISPR Protocols gRNA Design...CRISPR Cas9 gRNA plasmids from Dr. Rudolf Jaenisch's lab. CRISPR... CRISPR Jaenisch Lab CRISPR Plasmids Jaenisch...Design Tools CRISPR Blog Posts CRISPR-on is an RNA-guided transcriptional activator system with two components...: Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system...guide RNA (sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component ...activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression...
  25. Lentiviral Prep Service

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    ...Packaging Service page . Pooled CRISPR Libraries for Screening Use pooled CRISPR libraries to screen for genes...libraries 92385 Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set A Human CRISPR inhibition library...Root 92386 Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set B Human CRISPR inhibition library...RNA guide (gRNA) that harbors targeting sequences. Browse our repository for a list of gRNA plasmids targeting...genes. For more information on CRISPR plasmids and function, see our CRISPR plasmids resource page . Cas9...73179 Human gRNA pooled library in lentiCRISPRv2 Human sgRNA library in backbone lentiCRISPRv2 targeting...Service Lentivirus Browse In-Stock Lentivirus Pooled CRISPR Libraries Cas9 Pooled Barcoding Libraries Control...
  26. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...technologies like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID...check out this table.  All-in-one dual CRISPR/Cas12a (AIOD-CRISPR) assay – for rapid, ultrasensitive and...Resources on Addgene.org Get the basics about CRISPR in our CRISPR guide Browse our entire COVID-19 plasmid...detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-100 copies of...Lucia et al., 2020). Detecting coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic...the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification was able to detect as ... Cas13, upon finding complementarity between the gRNA and a given fluorescently labelled substrate, gets...
  27. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...Addgene.org Find all CRISPR plasmids and resources at Addgene Find CRISPR base editors Find CRISPR plasmids for...base. If you heard about CRISPR, one question probably comes to mind: can CRISPR be applied to fix the defective..., Y. Multiplex CRISPR-mediated base editing in rice. In Genome Engineering: The CRISPR-Cas Revolution,...Addgene blog Read all of our CRISPR 101 blog posts Download the CRISPR 101 eBook Read our blog posts ... to facilitate repair. Since the development of CRISPR-mediated base editing (BE), these types of repairs...et al., 2016 and Nishida et al., 2016) invented CRISPR-base editors by tethering cytosine deaminase with...adenine (A) to a guanine (G)? You may wonder if a CRISPR-base editor can perform this change. However, there...
  28. CRISPR 101: Epigenetics and Editing the Epigenome

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    Blog Post
    ...deacetylases. CRISPR and epigenetics As with many TALEN-based technologies, the advent of CRISPR has made .... For an up-to-date list of CRISPR epigenetic tools, check out our CRISPR Epigenetics resource. Transcriptional...potential advantage of CRISPR epigenetic tools is their persistence and inheritance. CRISPR activators and repressors... on Addgene.org Check out Our CRISPR Guide Pages Find Validated gRNAs to Target Your Genomic Locus of...modification at specific genomic loci is possible using CRISPR, and Addgene has a number of tools for this purpose...targeted to specific DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused...double strand breaks. Below you'll find some of the CRISPR-based epigenetic modifiers available from Addgene...
  29. 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 ...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...moves toward standardized thresholds for validating CRISPR experiments, many are turning to next-generation...
  30. Plan Your Experiment

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    Guide
    ...how to design your gRNA . Browse Plasmids: gRNAs Synthesize and clone desired gRNAs Once you have selected... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment Jump To: Plan Your Experiment... Select Expression System Design gRNA Publications Get Started CRISPR is a powerful system that enables...you started using CRISPR in your research. Although we will use the example of CRISPR/Cas9 in mammalian...Select Expression System To use CRISPR, you will need both Cas9 and a gRNA expressed in your target cells...express the CRISPR machinery. For more difficult cells (e.g. primary cells), viral delivery of CRISPR reagents... using CRISPR in mammalian cells. Some of the variables include: Species of Cas enzyme and gRNA Species...
  31. Pushing the Limits of DNA Assembly

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    Blog Post
    ...Resources on Addgene.org Browse Our CRISPR Resources Find Validated gRNAs Browse All Plasmids ...of base pairs. Cloning sgRNAs Generating constructs to produce sgRNA for CRISPR experiments can be laborious... plating, screening and generation of sgRNA. Generation of sgRNA libraries, with maximum complexity, is...constructs with multiple point mutations Cloning sgRNAs Production of Yeast Expression Cassettes Generating...synthetic oligo to generate a single-guide RNA (sgRNA). Using only one single-stranded oligo, instead ...to create a double-stranded oligo. Generating an sgRNA construct becomes as simple as combining your favorite...
  32. How to Deposit Your Plasmids with Addgene

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    Blog Post
    ... Encodes gRNA/shRNA A plasmid with one or more gRNA or shRNA sequences cloned into it. gRNAs are used...used for directing CRISPR/Cas9 to a targeted genomic location, while shRNAs are used to knockdown expression...gene contained within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert...
  33. CRISPR Guide

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    Guide
    ... of unique gRNAs. gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high ... Addgene Blog CRISPR topic page How to design your gRNA for CRISPR genome editing CRISPR 101: Homology...basic CRISPR mechanism Engineered CRISPR systems contain two components: a guide RNA ( gRNA or sgRNA ) and...Guide CRISPR Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large Edits Genome-Wide Screening...generating gRNAs makes CRISPR one of the most scalable genome editing technologies, making CRISPR perfect...Figure 7: Overview of CRISPR transposases Browse Plasmids: CRISPR transposases CRISPR Recombinases and Integrases...screening using CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries...
  34. Typing CRISPR Systems

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    Blog Post
    ...blog CRISPR 101: Cas9 vs. The Other Cas(s) CRISPR 101: RNA Editing with Cas13 Build your CRISPR vocabulary...Additional resources on Addgene.org CRISPR guide CRISPR plasmids CRISPR references ...Classification Structure Principles of CRISPR classification While CRISPR systems now seem neatly organized...versus class 2 There are two CRISPR classes: Class 1 and Class 2. Class 1 CRISPR systems use multi-Cas effectors... Type I CRISPR systems make up the majority of Class 1, and are the most common type of CRISPR overall...complex of the CRISPR types and is thought to be the common ancestor of all other CRISPR systems. Type ...immune functions that other CRISPR types do, due to a lack of canonical CRISPR features (Pinilla-Redondo...
  35. How to Keep a Lab Notebook for Bioinformatic Analyses

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    Blog Post
    ...Protocols for Your Next Experiment Find Our List of CRISPR gRNA Design Tools ...Save Time and Track Your Experiments Try out the CRISPR Software Matchmaker Additional Resources on Addgene.org...
  36. 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...linearized vector, the Cas9 enzyme along with a specific gRNA was used to cleave the 22kb vector. When followed...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 ...Wang A, Li K, Wang B, Jin S, Reiser M, Lockey RF. CRISPR/Cas9 nuclease cleavage combined with Gibson assembly...
  37. Bikard Lab - CRISPR Repression Collection

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    ...Lab - CRISPR Repression Collection Bikard Lab CRISPR Repression Plasmids You may also like... CRISPR Guide...Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The David Bikard Lab at the Institute Pasteur...a plasmid‐borne CRISPR array coding for guide RNAs (diamonds) interspaced with CRISPR repeat motifs (squares...From the Bikard Lab, CRISPR plasmid collection for modulating expression levels of two targets independently...Pasteur has deposited a collection of CRISPR arrays to modulate the expression levels of two targets independently... DNA-binding protein dCas9 is expressed along a CRISPR array that contains 20 bp "addresses", where dCas9...collection comprises 20 plasmids, each carrying two CRISPR guides. The first one binds to sfgfp with either...
  38. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

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    Blog Post
    ...Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease...Resources on Addgene.org Browse CRISPR plasmids Check out our CRISPR guide pages Find ready-to-use Cas9...intended target site. With the widespread adoption of CRISPR/Cas9 technology and rapid improvements in the field...delivery Epitope tagging of mammalian genes using CRISPR depends on successful delivery of three reagents...using custom synthetic two-part cr/tracrRNAs for CRISPR experiments (in collaboration with Integrated DNA...tagging is scalable to 96-well format. Off-the-shelf CRISPR reagents for the knock-in experiment were pre-assembled...csRNP knock-in experiments If your lab is new to CRISPR/Cas9 experiments, there is no need to worry at ...
  39. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR kits too...more of these kits, as there are many options! CRISPR CRISPR-Cas can harness the power of MoClo to expand... the most intuitive CRISPR MoClo application is to assemble multiple unique sgRNAs into a multi-TU targeting...including conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression...edit optimization, and more. While there are many CRISPR-Cas libraries and tools available for certain model...biologists, Addgene has not one, not two, but three CRISPR MoClo kits for plants that work for both monocots...MoClo can help you achieve it! More recently, a CRISPR MoClo kit termed Fragmid was developed for mammalian...
  40. Guide to Using Pooled Libraries

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    Guide
    ...Using CRISPR/Cas9 Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening Pooled CRISPR Libraries...protocol. Read our CRISPR guide for more information on genome-wide screening with CRISPR/Cas9 . Browse our...each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains a unique gene targeting...library from the Moffat lab has over 175,000 different gRNA-containing plasmids). Amplifying and Using the Library...of published selection mechanisms for lentiviral CRISPR libraries include: resistance to the anti-cancer... screen. The figure above is an outline of how a gRNA library is used. Below are the general steps to ...virus, make virus; this creates a pooled lentiviral CRISPR library. Apply pooled library to cells. DIVERGE...
  41. Plasmid Collections

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    Collection
    ...pooled libraries. Pooled Library Types cDNA shRNA CRISPR gRNAs Depositor Collections A modular set of plasmids...plasmids, kits, and pooled libraries. Find tools for CRISPR, fluorescent proteins, optogenetics, biosensors... Genome Engineering Genome Engineering Overview CRISPR Cre-lox TALENs Zinc Fingers Luminescence Fluorescent...view all collections. Depositor Collection Types CRISPR Fluorescent Proteins Protein Expression Synthetic...
  42. Twenty Years of Sharing: Expanding Accessibility

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    Blog Post
    ...Replication What is a Plasmid? How to Design Your gRNA for CRISPR Editing The Promoter Region - Let's Go! How...Educational Resource Top Five by Views Guides CRISPR Guide Lentiviral Guide AAV Guide Optogenetics Guide...primers to help users get started with tools like CRISPR, lentivirus, and optogenetics — and shared the ...Addgene blog Twenty Years of Addgene Sharing: CRISPRTwenty Years of Sharing: Addgene's Viral Vector Service...
  43. Worm Expression Resources

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    Collection
    ... individually. CRISPR Read Addgene's CRISPR Guide for a general introduction to CRISPR technology. Addgene's...Addgene's worm CRISPR resources include plasmids expressing Cas9, empty gRNA backbones, and constructs ... tag insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR plasmids from John Calarco's...genome editing in C. elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses purified...Browse plasmid tools, kits, and resources (such as CRISPR and synthetic biology collections) for use in C... feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific transgenic...contain tools for use in worm research. This includes CRISPR, RNAi, empty backbones, fluorescent proteins, and...
  44. Microbiology Resources

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    Collection
    ...Microbiology General CRISPR Resources Read our CRISPR Guide for a general introduction to CRISPR technology. Check...Check out our CRISPR reference page for CRISPR resources, such as depositor protocols and gRNA design software...software. Addgene yeast CRISPR resources Bacterial CRISPR Resources The CRISPR technology was developed...synthetic biology. CRISPR Resources : Find essential background information on using CRISPR in your lab, specific...Find CRISPR tools for use in genome editing of bacteria. CRISPathBrick Multiplex Plasmid : CRISPR expression... specific CRISPR tools for genome editing in bacteria, and more. Synthetic Biology : Browse our deposited...editing is not always straightforward. Read our CRISPR Methods for Bacterial Genome Engineering blog post...
  45. Zhang Lab CRISPR Page

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    Collection
    ...CRISPR Cas9 gRNA plasmids from the Zhang lab... Genome Engineering CRISPR Zhang Lab CRISPR Plasmids Zhang...Zhang Lab CRISPR Plasmids Available from Addgene Zhang Lab CRISPR Resources Tips for and FAQs about the ...Zhang Lab CRISPRs Browse All Zhang Lab Plasmids Zhang Lab Website You may also like... CRISPR Guide CRISPR...CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Jump to: SpCas9 Plasmids GeCKO SAM AAV - In vivo Delivery...phages and plasmids. CRISPR loci in microbial hosts contain a combination of CRISPR-associated (Cas) genes...specificity of the CRISPR-mediated nucleic acid cleavage. We have harnessed the type II CRISPR nuclease system...
  46. Bacterial Expression Systems

    Type
    Collection
    ... list of our bacterial CRISPR plasmids, see here . pgRNA-bacteria 44251 CRISPR Stanley Qi Expression of...Purpose pCas9 42876 CRISPR Luciano Marraffini Bacterial expression of Cas9 nuclease gRNA. For a more comprehensive...list of our bacterial CRISPR plasmids, see here . pwtCas9-bacteria 44250 CRISPR Stanley Qi Anhydrotetracycline...can be combined with expression of a gRNA from a plasmid like pgRNA-bacteria to knock down endogenous gene...alter bacterial genomes. From recombineering to CRISPR , this collection contains a variety of tools to... of customizable guide RNA (gRNA) for bacterial gene disruption or knockdown. For a more comprehensive...comprehensive list of our bacterial CRISPR plasmids, see here . pGRG Series Various Plasmids Tn7 Transposition Nancy...
  47. Zebrafish Plasmid Collection

    Type
    Collection
    ...Collection CRISPR Please refer to Addgene's CRISPR Guide for a general introduction to CRISPR technology... zebrafish CRISPR resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones....examples of useful zebrafish CRISPR resources can be found below: UFlip CRISPR targeted integration vectors...for Targeted Integration Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression...Joung Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9 Vector...modified CRISPR-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference...for the laboratory use of zebrafish. CRISPRz - A database of CRISPR/Cas9 target sequences that have been...
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