...lentiviral expression vector, which contains an sgRNA insert site and a reporter for tracing the engineered...editing tool on the immune system, the group inserted sgRNAs targeting various innate and adaptive immune genes...

...Cas9 from Streptococcus pyogenes with an invariant sgRNA scaffold and cloning sites for inserting the guide...pyogenes CRISPR chimeric RNA element with customizable sgRNA from the U6 promoter and puromycin resistance from...

...PU6::unc-119_sgRNA : unc-119targeting sgRNA. 46170 PU6::klp-12_sgRNA : klp-12targeting sgRNA. Please note.... elegans . The protocol for customizing the sgRNA expression plasmid can be located in the Materials...

...46,354 BARBEKO sgRNA Library 174163 Knockout, Base Editing Human Wei 3rd 3 53,502 sgRNA Pool for Streptococcus...131625 Inhibition E. coli Bikard N/A ∼5 21,417 Bison sgRNA Library 169942 Knockout Human Ebert 3rd 4 2,852 ...Fly Perrimon 3rd 8 6 6 7,956 17,827 59,406 Dual sgRNA CRISPRi Libraries 187246 — 187248 1000000192 Inhibition...Inhibition Human Weissman 3rd 6 Varies Dual sgRNA CRISPRa Libraries 187249 — 187251 1000000193 Activation...3rd 2 37,722 Green monkey (Chlorocebus sabaeus) sgRNA library 178284 Knockout Green Monkey Doench and ...Knockout Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library 140633 Knockout Human Wei 3rd 3 58,... Mouse Knouse 3rd ~5 71,878 Mouse messenger-RBP sgRNA Library 169082 Knockout Mouse Turner N/A 10 13,500...

... an sgRNA uder the Arabidopsis U6 promoter. 46966 pICH86966::AtU6p::sgRNA_PDS : Expresses an sgRNA targeting...manuscript for the protocol on assembling customised sgRNA expressing plasmids. Questions regarding the protocol...module. 49772 pAGM4723::AtU6p::sgRNA2-2x35S-5'UTR::Cas9::NOST-AtU6p::sgRNA1 : Level 2 construct....

...expression, human optimized 48673 M-NM-sgRNA Mammalian U6-driven sgRNA (NMm1) targeting GTCCCCTCCACCCCACAGTG...expression, human optimized 48672 M-ST1-sgRNA Mammalian U6-driven sgRNA (STm1) targeting GTCCCCTCCACCCCACAGTG...III at five protospacers 48671 M-SP-sgRNA Mammalian U6-driven sgRNA (SPm) targeting GTCCCCTCCACCCCACAGTG...

...a two viral vectors system: one vector for Cas9/sgRNA and one vector for your specific DNA template. Using...by John Doench CRISPR-Cas9: Tips for Optimizing sgRNA Activity by John Doench All CRISPR Posts  ...

...PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco SP6-sgRNA-scaffold ...pyogenes (PAM = NGG) pDD162 (Peft-3::Cas9 + Empty sgRNA) R07E5.16 U6 yes, cut Goldstein DR274 T7 BsaI In...none, need Cas9 plasmid Sternberg L4440_BioBrick-sgRNA T7 BbsI, BsaI Ingestion by C. elegans none, need...

...Genome Editing CRISPR-Cas9: Tips for Optimizing sgRNA Activity Lentiviral CRISPR Libraries Enable Genome-Scale...

...those generated by multiplexed delivery of multiple sgRNAs. Since multiplexing is commonly used to create ...helps the researcher see which of the multiplexed sgRNA was involved in a particular edit, and which type...

... the Zhang lab created an all-in-one SaCas9 and sgRNA construct using the liver-specific serotype AAV8... a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs mediate genome modification, but they...

... the BFP variant, a BE4am-Cas9 base editor, and sgRNA vectors for the BFP-to-GFP conversion and to target...cytidine base editor. The lab then introduced an sgRNA targeting the GFPACG to the cells. As expected, ...

...trace files to the TIDE tool online along with your sgRNA sequence. The TIDE software will generate a graph...along with the estimated editing frequency of your sgRNA-Cas9 combo. Pro tip: When designing your PCR, ensure...

...vectors for the delivery of the synthetic guide RNA (sgRNA). Each of the Cas9 vectors contains the Cas9 coding...with and without a C-terminal EGFP fusion. For the sgRNA, vector pMB60 allows in vitro production of RNA ...

...lab used single-AAV delivery of Nme2Cas9 and the sgRNA to demonstrate efficient editing in adult mice and...

... expression. They replaced the tetraloop of the sgRNA scaffold with a com aptamer which binds an aptamer...

... 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 ...

...of 5%. Your experiment involves making a large sgRNA library to thousands of genes and targeting and ...sequencing millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to...

...References: 1. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize off-target...

...is described in Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation. Doench...Doench & David Root and are described in Optimized sgRNA design to maximize activity and minimize off-target...