QIAquick Gel Extraction Kit
Company: QIAGEN
Catalog#: 28704
Product evaluation:
Protocol for Construction of a Tunable CRISPR Interference (tCRISPRi) Strain for Escherichia coli
Author:
Date:
2017-10-05
[Abstract]  We present a protocol for construction of tunable CRISPR interference (tCRISPRi) strains for Escherichia coli. The tCRISPRi system alleviates most of the known problems of plasmid-based expression methods, and can be immediately used to construct libraries of sgRNAs that can complement the Keio collection by targeting both essential and nonessential genes. Most importantly from a practical perspective, construction of tCRISPRi to target a new gene requires only one-step oligo ...

Dense sgRNA Library Construction Using a Molecular Chipper Approach
Author:
Date:
2017-06-20
[Abstract]  Genetic screens using single-guide-RNA (sgRNA) libraries and CRISPR technology have been powerful to identify genetic regulators for both coding and noncoding regions of the genome. Interrogating functional elements in noncoding regions requires sgRNA libraries that are densely covering, and ideally inexpensive, easy to implement and flexible for customization. We present a Molecular Chipper protocol for generating dense sgRNA libraries from genomic regions of interest. This approach utilizes a ...

Targeted Nucleotide Substitution in Mammalian Cell by Target-AID
Author:
Date:
2017-06-05
[Abstract]  Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems have been applied to various type of cells as powerful genome editing tools. By using activation-induced cytidine deaminase (AID) in place of the nuclease activity of the CRISPR/Cas9 system, we have developed a genome editing tool for targeted nucleotide substitution (C to T or G to A) without donor DNA template (Figure 1; Nishida et al., ...

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