We narrowed to 181 results for: grna
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TypeBlog PostPublishedJuly 13, 2023, 1:15 p.m....Addgene, including cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries at Addgene...
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The AAVantages of AAV in CRISPR Screens
TypeBlog PostPublishedFeb. 4, 2025, 2:15 p.m....dedicated to expressing Cas9, while the other houses the gRNA and scaffold. If you don’t feel like doubling the... -
CRISPR Antimicrobials
TypeBlog PostPublishedMay 3, 2016, 2:30 p.m....Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog PostPublishedNov. 21, 2024, 10:47 p.m....Replication What is a Plasmid? How to Design Your gRNA for CRISPR Editing The Promoter Region - Let's Go... -
CRISPR-mediated Plant Base Editors
TypeBlog PostPublishedJan. 3, 2019, 1:35 p.m....expression system. We use a single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog PostPublishedMay 5, 2020, 1:15 p.m.... 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 ... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....can bind loci in the genome that are defined by a gRNA. Since Cas9 is a protein, a fusion with GFP allows... -
Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods
TypeBlog PostPublishedMarch 1, 2016, 3:30 p.m....linearized vector, the Cas9 enzyme along with a specific gRNA was used to cleave the 22kb vector. When followed... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog PostPublishedJune 24, 2020, 5:45 p.m....Check out Our CRISPR Guide Pages Find Validated gRNAs to Target Your Genomic Locus of Interest Browse ... -
Hot Plasmids: Spring 2025
TypeBlog PostPublishedJune 3, 2025, 1:15 p.m....scarless insertion of the payload (teal). The Cas9 gRNA targeting guide and the transposon RNA homology ... -
Sequencing Options for CRISPR Genotyping
TypeBlog PostPublishedOct. 4, 2016, 2:30 p.m....position of the mismatch along the length of the gRNA given the stricter sequence requirement at the terminal...many software tools predict off-target effects of sgRNAs using computational methods. They identify possible...mismatches based on the sequences of the genome and sgRNA. This is a good starting point for most researchers... -
Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
TypeBlog PostPublishedJune 28, 2018, 3:01 p.m....genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA. J Biotechnol 241:...complexed with sgRNA. Direct comparison of Cas9 RNPs prepared with an in vitro-transcribed (IVT) sgRNA versus ...cumbersome in vitro synthesis of single guide RNAs (sgRNAs), are time-consuming, have been typically optimised... the in vitro-transcribed single guide RNA (IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not...construct plasmid vectors or synthesize/purify IVT sgRNA in the lab. Figure 2: Cas9 RNP complexed with 2... -
Plasmids 101: Modular Cloning Applications and Kits
TypeBlog PostPublishedMay 14, 2024, 1:15 p.m....repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized CRISPR kits too...assemble multiple unique sgRNAs into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This ...for the multiplexing of sgRNAs against unique genomic targets or multiple sgRNAs against the same target...target to ensure efficient cutting. Beyond sgRNAs, MoClo can also be used to test a variety of Cas enzymes... -
Typing CRISPR Systems
TypeBlog PostPublishedMarch 18, 2025, 1:15 p.m....Cas12 is unique because it can process multiple gRNAs under a single promoter, allowing for easy multiplexing... -
28 Hot Plasmid Technologies from 2015
TypeBlog PostPublishedDec. 23, 2015, 3:30 p.m....targeted to their endogenous loci using validated gRNAs from Addgene and well designed homology directed...fused to an sgRNA used to direct Cas9 to a specific genomic locus. When fused to the sgRNA and co-expressed...dCas9 – the dCas9, sgRNA/RNA cargo complex localizes to the sequence targeted by the sgRNA. This allows scientists...machinery that brings the sgRNA/cargo RNA to the appropriate DNA locus. The sgRNA/RNA fusions retained their...construct. The sgRNA targeting Gaussia luciferase can be easily replaced with an sgRNA targeting your ...specific recruitment of multiple copies of VP64 to the sgRNA-targeted gene, resulting in increased activation...SV40, SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive ... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog PostPublishedOct. 4, 2018, 12:44 p.m....CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per gene...contains two sets of sgRNAs; A, the top 3 ranked sgRNAs, and B, the next 3 ranked sgRNAs. Dolcetto was able...containing only 3 sgRNAs (setA) also outperformed CRISPRi libraries with 10 sgRNAs in the sgRNA metrics, confirming...2014 to predict sgRNA activity. In this study scientists examined approximately 1,900 sgRNAs and identified...subsampling analysis of the sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR...CRISPR library with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello... CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While... -
Prime Editing: Adding Precision and Flexibility to CRISPR Editing
TypeBlog PostPublishedJan. 13, 2025, 9:49 p.m....engineered pegRNA” or epegRNA. Although the addition of the RNA pseudoknot sometimes altered epegRNA expression...does just this by including an additional sgRNA. Using this sgRNA, the prime editor nicks the unedited strand...4.0 license. epegRNAs: Structured RNAs with improved stability Traditional sgRNAs are protected from...degraded, the truncated pegRNA can still guide Cas9 and compete with full pegRNAs for access to the target... compared to PEmax with normal pegRNAs, but combining PE7 with epegRNAs resulted in similar or reduced... ~ 3′ RNA pseudoknot (epegRNA) Improve pegRNA stability ~ ~ ~ ~ ~ ~ ✖ ...? First, an engineered prime editing guide RNA (pegRNA) that both specifies the target site and contains... -
Hot Plasmids Spring 2024
TypeBlog PostPublishedMay 23, 2024, 1:15 p.m....engineered pegRNAs (epegRNAs), which add a structural motif to stabilize the 3’ end of the pegRNA. As research...The team designed sgRNAs that include an internal barcode and separated multiple sgRNAs with tRNAs, which...BFP2. Figure 1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using...lentiviral integration, producing a second copy of the sgRNAs. Image reused from Walton et al. 2024 under a CC-BY-NC-ND...occurs on the RNA polymerase III transcript, the sgRNAs were encoded on the lentiviral minus strand in ...of host cell proteins, and modifications to the pegRNA. The Britt Adamson Lab recently used a CRISPRi ...to La binding and stabilizing the 3’ end of the pegRNA, which is not well protected by Cas9. This is similar... -
Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens
TypeBlog PostPublishedFeb. 24, 2015, 7:50 p.m.... 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...further optimize those now. They are also developing sgRNAs that will enable even more fine-tuning of gene ... -
A Needle in a Base-Stack: Cas9 Structural Biology
TypeBlog PostPublishedJune 4, 2024, 1:15 p.m....Once supplied with a guide RNA — whether it’s an sgRNA or a crRNA:tracrRNA duplex — Cas9’s two lobes move...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue. The REC lobe binds tightly to ...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue. DNA sequence recognition Once ...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue; target DNA strand, light gray; non-target... and marks the transition back to a DNA duplex. sgRNA, dark blue; target DNA strand, light gray; non-target...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue; target DNA strand, light gray; non-target...