Why should I buy the Guide-it CRISPR Genome-Wide sgRNA Library System?

-sgRNAs are designed using the Brunello algorithm High confidence that a high percentage of the individual sgRNAs are active and specifically target the expected gene. The high sgRNA activity also supports the use of a smaller number of sgRNAs/gene. -Targets >19,000 genes The user can perform an unbiased analysis by screening against the entire human genome. -Includes four different guides [...]

What is a typical timeline for a genome-wide CRISPR screen using a pooled lentiviral sgRNA library?

Genome-wide CRISPR screens using pooled lentiviral sgRNA libraries typically take several months to complete. NOTE: The workflow below does not include the time required to produce the sgRNA library. Our kit provides the researcher with a high-quality, validated library for expediting the process.Gene editing [...]

Is there any screening data for the Guide-it CRISPR Genome-Wide sgRNA Library System?

Yes. Click on the link to see data generated in a screen for genes involved in resistance to the purine analog 6-thioguanine (6-TG). https://www.takarabio.com/learning-centers/gene-function/gene-editing/genome-wide-screenin... Technical note: Performing a phenotypic screen using the Guide-it CRISPR Genome-Wide sgRNA Library SystemGene editing [...]

Should I be worried about false positives or off-target effects when screening using the Guide-it CRISPR Genome-Wide sgRNA Library System?

False positives from a screen can result from a guide RNA targeting an off-target sequence that is unrelated to the sequence for which it was designed. We have significantly negated the risk of this happening with the design of the library and the optimization of the screening process. This library contains sgRNAs from the Brunello library which were chosen using algorithms that maximize [...]

How much DNA do I need to isolate prior to performing NGS?

Although you only need a small amount of DNA for sequencing, we recommend that you isolate DNA from 100–200 million cells for pooled screens. This typically amounts to 300–500 µg of genomic DNA. The reason for this is that it is essential to generate an NGS library that is representative of the diversity of the edited cells (and hence sgRNAs) still in the population after treatment. Purifying [...]