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We narrowed to 176 results for: grna design

Showing: 121 - 160 of 176 results

  1. Microbiology Resources

    Type
    Collection
    ...CRISPR resources, such as depositor protocols and gRNA design software. Addgene yeast CRISPR resources Bacterial...recombinant protein purification and cell-free circuit design in E. coli - Freemont Lab MoClo-YTK : Assembly ...

  2. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...target DNA sequence as directed by the guide RNA (gRNA), while the CRY2PHR-activator fusion floats freely...shown in the figure below. Their most successful design utilized Magnet photoswitchable proteins derived...occurs. Bubeck et al. expressed a telomere-targeting gRNA and a dCas9 fusion protein with three red fluorescent...Photocleavable ssDNA oligonucleotides Cas9, photocaged-gRNA Gene editing Manna et al., 2019   NPPOC-caged ..., 2020   NPOM-caged nucleobases Cas9, photocaged-gRNA Gene editing For more details, check out this...

  3. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ...target DNA, recruited by either the ωRNA (TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.com...different applications require careful selection or design to ensure reliable readout and avoid biases. The...ex vivo HSC expansion cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures...only at excitatory synapses; however, its modular design allows components to be swapped out, potentially... Huang, H., et al. (2025). ATLAS: a rationally designed anterograde transsynaptic tracer. Nature Methods...

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

    Type
    Blog Post
    ...smaller CRISPR-Cas proteins such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter...this post, you’ve got the knowledge to be able to design a construct for the successful and specific expression...

  6. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...Origin of Replication What is a Plasmid? How to Design Your gRNA for CRISPR Editing The Promoter Region - Let's...Electrophoresis Bacterial Transformation How to Design a Primer Creating Bacterial Glycerol Stocks Inoculating...Analysis Videos What is a Plasmid? How to Design Primers for PCR Agarose Gel Electrophoresis Inoculating...all 101s on a particular topic into beautifully designed and easy-to-reference eBooks. Figure 3: Global...changes, like using alt-text, captions, and inclusive design in our digital content. We have implemented proactive...

  7. CRISPR Antimicrobials

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    Blog Post
    ...Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ...currently undergoing a renaissance. Yosef et al. designed a two-phage CRISPR system to resensitize antibiotic-resistant...Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR design for next-generation antimicrobials.” Genome Biol...

  8. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    ...Cas9. With many PAMs available, a large number of gRNAs could be screened for the most efficient editing...final base editor that was administered to KJ was designated NGC-ABE8e-V106W. The name is hardly flashy, but...different mutations in different patients. While these gRNAs would still need to be optimized and assessed for...the best when paired with the highest-efficiency sgRNA. The PAM sequence near this site is AGC, so in a...

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

    Type
    Blog Post
    ... Both Snapgene and NEB have tools that help you design primers for PCR amplification of fragments to incorporate...demonstration of how to use Snapgene’s program to design primers for Gibson Assembly. For a simple example...without scars as depicted below. First, you need to design primers to amplify the two fragments while also...assembly, long primers for each fragment must be designed and ordered, and are specific for each fragment...linearized vector, the Cas9 enzyme along with a specific gRNA was used to cleave the 22kb vector. When followed...ratios of product to insert, and tools for primer design, so it’s always best to check in with the instructions...

  11. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...please improve your plasmid maps!" After thoughtful design, vetting, and tweaking, we’re excited to announce...commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display, you...

  12. Hot Plasmids: Spring 2025

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    Blog Post
    ...Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren Looger and Kaspar Podgorski...Aggarwal et al., 2025).In this latest work, the team designed and screened a large library of iGluSnFR3 mutants... HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's what makes these... stable cell line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked... plasmids for flexibility and speed, so you can design, test, and innovate faster than ever. Plus, we'...scarless insertion of the payload (teal). The Cas9 gRNA targeting guide and the transposon RNA homology ...

  13. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...introduced through non-homologous end joining (NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component...edit to be made when multiple As are present in the gRNA spacer, as ADAR will preferentially edit an adenine...various spacer lengths from 30-84 nucleotides, and designated this system REPAIRv1. Using next-generation sequencing...

  14. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... This vastly increases the possibilities when designing screens. Each serotype has its own tropism, or...dedicated to expressing Cas9, while the other houses the gRNA and scaffold. If you don’t feel like doubling the...Adames, A. C., Saur, D., & Rad, R. (2022). Tutorial: design and execution of CRISPR in vivo screens. Nature...

  15. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ... Gateway compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors...needed for replication in the host cell and are designed for a user to insert a gene of interest. They ...start codon for N-terminal tags. And when you are designing your plasmid you should keep your gene "in...Plasmids and Resources collection, and browse our Empty gRNA Expression Vectors page Cre-lox Site-specific recombination...selectable marker pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP - For retroviral delivery of one sgRNA pMKO.1 GFP - Retroviral...- Deliver sgRNA and Cas9 into plants by Agrobacterium-mediated transformation Dual-sgRNA_hU6-mU6 - Lentiviral...Zeocin®/Bleo Mammalian lenti sgRNA(MS2)_zeo backbone - 3rd gen lenti sgRNA cloning backbone with MS2 loops...

  17. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

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    Blog Post
    ... systems, you will have to design your crRNAs using other web-based design tools (for C-terminus tagging...knock-in. Another good design tool can be found at Desktop Genetics .For ssODN design, remember to incorporate...mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA. J Biotechnol...bioinformatics tool for guide RNA and donor DNA design, and implemented our optimised protocols into a...genome browser: https://www.ensembl.org/index.html Design crRNA/ssODN: Using Ensembl Transcript ID(s) as ...Dewari et al. 2018). Using the first crRNA we designed for ~250 genes, we have had about 30% tagging ... in a couple of weeks by combining our “TAG-IN” design tool with the csRNP method. We anticipate that ...

  18. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ... system developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target DNA without cleavage until...

  19. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...good reason. It is guided by an RNA molecule, the gRNA, to a complementary site in the genome where it ...in one direction. Cas3 is thus not helpful for designer genome edits, but it is effective at wiping out...

  20. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...many MoClo kits are designed with specific model organisms in mind, others are designed for flexibility. ...Synthetic biology in bacteria MoClo is commonly used to design TUs and multi-TU constructs for synthetic biology.... phaffii, then the MoClo YTK Kit specifically designed for S. cerevisiae is at your service (Lee et al...repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR kits too... the Multi-Kingdom (MK) Golden Gate system was designed to work in fungi, bacteria, protists, plants, ...E. Coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology.” ACS Synthetic...Mammalian Modular Assembly Toolkit for the Rapid Design and Production of Diverse Expression Vectors.” ...

  21. dTAG - You're it!

    Type
    Blog Post
    ...Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous...CRISPR plasmids for protein tagging Find validated gRNAs ...

  22. Zebrafish Plasmid Collection

    Type
    Collection
    ... selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of useful zebrafish CRISPR...promoters, six fluorophores, and empty vectors designed to streamline the production of transgenic zebrafish...

  23. Hot Plasmids: Fall 2025

    Type
    Blog Post
    ..., 2025). These new sensors use quite different designs, leading to unique advantages. While most red GECIs...two-photon microscopy, and fiber photometry. Plus, both designs offer promising templates for future improvements...transcription and translation of chromoprotein and gRNA plasmids. Nuclease-dead Cas9 (dCas9) protein is ...

  24. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...one temperature. Two Cas12a-crRNA complexes are designed, each binding to one strand of the reverse-transcribed... Cas13, upon finding complementarity between the gRNA and a given fluorescently labelled substrate, gets... E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds to the target sequences, Cas12a activity...Fluorescently labelled dCas9 complexes containing sgRNA that have no mismatches bind to the immobilised ...

  25. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...expression system. We use a single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers... SpCas9-VRER TadA-TadA* NGCG 8  Tips for designing base editing experiments in plants Choose a base...

  26. CRISPR 101: Epigenetics and Editing the Epigenome

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    Blog Post
    ...certainly not the only CRISPR-based technology designed to alter gene expression. Fusing dCas9 to a transcriptional...Check out Our CRISPR Guide Pages Find Validated gRNAs to Target Your Genomic Locus of Interest Browse ...

  27. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ... Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching... the DNA strand that does not base pair with the gRNA. Base editors target this “non-target strand.” To... predicting CGBE editing outcomes and assisting sgRNA selection is available at http://www.sunlab.fun:... in different cell types, available at www.crisprbehive.design. CGBEs fused to pioneer factors to improve...

  28. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...CRISPR library with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello...2018). These libraries utilize optimized sgRNAs that were designed based on the rules identified in Doench...gene and 1,000 non-coding control sgRNAs. This library was designed to improve on-target activity while...confirming again that sgRNA design plays a significant role in library efficiency. Try the Dolcetto library...uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA aptamers.... Doench, John G., et al. "Rational design of highly active sgRNAs for CRISPR-Cas9–mediated gene inactivation...MC4262738. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize off-target...

  29. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...stepwise protocol on designing and using RNA scaffolds: Delebecque, C. J. et al. Designing and using RNA scaffolds... of transcriptional regulator or the number of designed scaffold binding sites. With hundreds of characterized...tunability of transcription is possible. From a design perspective, this is appealing since the RNA scaffold...pairing, allowing for rational, straightforward design. Since the technique relies on base pairing, it...and multidirectional multiplex control via scRNA design is readily apparent in metabolic engineering. The...targeted site. Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional and tunable...activity coupled with a customizable single guide RNA (sgRNA) that bears complementarity to a 20bp target sequence...

  30. Uncovering Ancestral Bias in CRISPR Research

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    Blog Post
    ...they designed a tool that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having... guides used in the screens they analyzed were designed to have a perfect match to the reference genome...Brown from Broad's Genetic Perturbation Platform to design a new library that is agnostic to ancestry which...CRISpick algorithm to prevent guides from being designed that target regions with high genomic variability...actually an artifact introduced by bias within the design of the experiment or dataset itself. It is important...scientists to be aware of and actively use tools designed to help reduce or eliminate bias, just like the...indicating loci where SNPs reside on sgRNA target sequences for sgRNAs that target ancestry-associated genes...

  31. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ... endogenous loci using validated gRNAs from Addgene and well designed homology directed repair templates...cell events Three years ago, Brian Kuhlman’s lab designed photoswitchable dimers by fusing the SsrA peptide...the authors recently used computational protein design, phage display and high throughput binding assays...gene is inserted. It is important that primers be designed to minimize the possibility of dimer formation...platform for imaging and flow cytometry applications Designing customized fluorescent reporters can be challenging...preposterous. Luckily, the Pierre Neveu laboratory designed a new iterative, chaining-based cloning method...the field of all-optical electrophysiology is designing the perfect voltage indicator and optogenetic ...

  32. A Tour of Addgene's Most Popular Pooled Libraries

    Type
    Blog Post
    ...the only game in town! Pooled libraries can be designed for all kinds of different high-throughput experiments...elements using golden gate cloning. This toolkit is designed to be used across many species of bacteria. Surface...Libraries has tips on using these libraries and designing screens with them. All pooled libraries are available...CRISPR screening libraries supply a collection of sgRNAs, but they may or may not include CRISPR machinery... CRISPR libraries In CRISPR knockout screening, sgRNAs guide Cas9 to cut targeted genes, creating mutations...screening are similar to those for knockout screens: sgRNAs to guide Cas9 to targeted genes. In these applications...position, and sequence features in the selection of sgRNAs. Other CRISPR libraries As CRISPR technologies ...

  33. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind. “We had not managed...from the structure to guide and improve activator design,” she said. “Before it was a little like tapping...instead of at the protein’s C terminus. With that new design in place, Konermann immediately saw an order of... activator (CRISPRa) sgRNA library uses the sunCas9 system and contains 10 sgRNAs for each transcription...nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop...gain-of-function screening when combined with a compact pooled sgRNA library. Our SAM-mediated screens exhibited a high...transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each transcription start site in those 15,977...

  34. Hot Plasmids: Fall 2024

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    Blog Post
    ...promising tool for future structure-guided drug design and studies of protein ubiquitination and degradation...CRBNmidi plasmid here!   Kroupova, A. et al. (2024). Design of a Cereblon construct for crystallographic and...specifically for C. elegans (Cao et al., 2024). The team designed coding sequences for mScarlet3 and mScarlet-I3...recombination methods). By using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ ...Using an inducible Cas9 expression and removing the sgRNA plasmid after identifying successful knockouts limits... KanR. This strain is then transformed with the sgRNA cassette on a plasmid expressing mCherry and resistance... of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain carrying Cas9 to...

  35. Hot Plasmids and Viral Preps - September 2021

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    Blog Post
    ...without end-point sequencing assays.  This tool was designed by putting together the coding sequence of a mCherry...MYC-CRISPR library targets E-boxes genome-wide. It was designed based on MYC-ChIP-seq data from several MYC-dependent...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons... cell lines. The Turner Lab Human messenger-RBP sgRNA Library targets human mRNA binding proteins with...operons in other pneumococcal strains. The BARBEKO sgRNA library uses CRISPR cytosine base editors to disrupt...

  36. Tetracycline Inducible Expression

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    Collection
    ...promoters (sometimes generally called pTet) have been designed by combining tet O sequences with elements like...as much flexibility and control as a dual vector design, but it is easier to implement and ensures that...Lentiviral Tet-On plasmid for easier cloning and design of inducible shRNA expression, with puromycin selection...104321 tet-pLKO-sgRNA-puro Lentiviral Tet-On plasmid for inducible expression of sgRNA TetR H1-2O2 Nathanael...

  37. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    ... Vectors The vectors pK-TnpB1 and pK-TnpB2 are designed for plant genome editing using TnpB. The main ...step-by-step guide for using a Dra2TnpB. Step 1: Design the 20-nt Guide Sequence Identify a TAM site (...schematic of pKb-TnpB1 and pKb-TnpB2, and guide design rules.   Step 3: Vector Digestion Digest either...single guide RNA (sgRNA) used in Cas systems. The structure is also reminiscent of sgRNAs, with reRNA containing...

  38. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...domain (FokI). Zinc finger nucleases were first designed in 2005. The DNA-binding domain can be engineered...editors, new amino acid recognition sites had to be designed each time we wanted to target new genomic sites...chasing an anti-villain geneticist named Zobrist who designs a virus that can potentially kill one-third of ... in genome editing is prime editing, which was designed in 2019. It’s based on a prime editor which is... Using an elongated template + guide RNA called pegRNA (prime editing RNA), the prime editor can install...

  39. Plan Your Experiment

    Type
    Guide
    ...used for gRNA design. Read more about how to design your gRNA . Synthesize and Clone Desired gRNAs Once your... proper gRNA design. While there are some cases where it makes sense to manually design a gRNA, in most...most cases gRNA design software is better and more efficient at designing potential gRNAs for you to choose...selecting the edit type, designing and cloning a gRNA, delivering Cas and the gRNA, and validating the edit... system for Cas9 and the guide RNA (gRNA), the design of the gRNA sequence, and finally how to validate...target and designed your gRNA(s), the next step is to choose how to express your Cas enzyme and gRNA(s) in ... multiple gRNAs is a good idea for most CRISPR experiments, testing multiple pegRNA designs is particularly...
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