Titlebook: CRISPR Guide RNA Design; Methods and Protocol Tudor A. Fulga,David J. H. F. Knapp,Quentin R. V. Book 2021 Springer Science+Business Media,

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Controlling the Activity of CRISPR Transcriptional Regulators with Inducible sgRNAsthe presence of a user defined inducer. In this chapter, I cover the know-how relating to the design and synthesis of iSBH-sgRNAs, as well as the implementation in mammalian cells of inducible CRISPR-TR strategies based on this technology.

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Gene Manipulation Using Fusion Guide RNAs for Cas9 and Cas12as of the CRISPR-Cas system. Recently, we have developed fusion guide RNAs (fgRNAs) for orthogonal gene manipulation using Cas9 and Cas12a. Here, we describe the methods for designing and generating fgRNAs-expression constructs to achieve multiplex genome editing and gene manipulation in human cells.

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The Thin Filament in Insect Flight Muscleprovide a detailed protocol for use of CRISPR-Cpf1 and chemically modified guide RNAs in cell lines, outlining the steps from designing guide RNAs to a target gene of interest, delivery and expression in cells, and analysis of gene editing events.

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The Thin Filament in Insect Flight Muscle active sgRNA. The benefits of our approach lie in the stringent purification of the DNA-targeting 20-mer, the reduced synthesis of the constant 79-mer each time a new sgRNA is required, and the rapid access it provides to custom libraries of sgRNAs.

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Noch einmal: Geschichte in Schichtgesteinen,identify off-target cleavage events throughout an entire genome. Together, these techniques can be used to assess the reliability of experimental models generated using CRISPR/Cas9 as well as the safety of therapeutics employing this technology.

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