... (PAM = NGG) 47549 pDD162 (Peft-3::Cas9 + Empty sgRNA) R07E5.16 U6 yes, cut Goldstein 42250 DR274 T7 BsaI...transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold SP6 AflII In vitro transcription none, ...need Cas9 plasmid Sternberg 177783 L4440_BioBrick-sgRNA T7 BbsI, BsaI Ingestion by C. elegans none, need...transcription none, need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9...

...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...

...shRNA sequence specific to your gene’s mRNA or a sgRNA targeting an exon in your gene of interest, for ...with many applications. These libraries include sgRNAs targeting the entire genome, overexpression of ...

...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  ...

...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...

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

... 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...

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

... 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...

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

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

...60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS... Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang...ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression...expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks 60877 pMAZe...

...regulator. The lab expressed both dCas9 and the ctrA sgRNA and saw that the levels of ctrA mRNA decreased after...

...sequential infection first of Cas9, then with the sgRNAs, and can result in a higher titer due to the smaller...

... plate, co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100bp ... a typical size (~ 5kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example...of a typical size (~5kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good...

...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...