What is HDR for Gene Editing?

HDR stands for ""Homology-directed repair"", a method of providing intended editing by exogenous DNA to be integrated into the host genome.Gene editing [...]

What additional kits are needed for preparing NGS libraries downstream of screening?

The Guide-it CRISPR Genome-Wide sgRNA Library NGS Analysis Kit (Cat. # 632647) enables preparation of Illumina-ready NGS libraries from cell populations screened using the Guide-it CRISPR Genome-Wide sgRNA Library System (Cat. # 632646), allowing for identification of sgRNA sequences and corresponding gene knockouts that have increased or decreased in frequency under the conditions of the [...]

What is the advantage of having an mCherry reporter gene in the sgRNA vector?

The red fluorescent protein mCherry is expressed from the same vector that expresses each sgRNA and is included to simplify the process of determining the optimal MOI for the screen. The aim is to obtain a transduction efficiency of 30–40% in your target cells, which ensures that most transduced cells contain only a single sgRNA construct. For the calculation of transduction efficiency, a [...]

Where can I find all the vector maps for the Guide-it CRISPR Genome-Wide sgRNA Library System?

Below are links to the sequence and map files for the vectors used in the Guide-it CRISPR Genome-Wide sgRNA Library System. Cas9 vector - map file: https://www.takarabio.com/assets/documents/Vector%20Documents/pLVXS-EF1a-Cas9-PGK-Puro%20V... Cas9 vector - genbank sequence file: https://www.takarabio.com/assets/documents/Vector%20Documents/pLVXS-EF1a-Cas9-PGK-Puro%20V... sgRNA Library vector - [...]

What are the advantages of your lentiviral packaging system, and how is it different from other libraries?

Our library is supplied lyophilized in our Single Shots format with our fourth-generation lentiviral packaging system, designed to produce high titers (up to 108 IFU/ml) without the need for optimization or post-production processes, such as concentration. A high-titer virus is essential for library screening and is produced by simply reconstituting this mixture with sterile water and adding it [...]