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PCRISPR007

Sigma-Aldrich

Human CRISPR Brunello Knockout Library

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NACRES:
NE.02

packaging

pkg of 5 vials (5x200µL aliquots )

concentration

≥5x108 VP/ml (via p24 Assay)

application(s)

CRISPR

shipped in

dry ice

storage temp.

−70°C

General description

The human CRISPR ′Brunello′ pooled library is designed using optimized metrics, as published by, Doench et al. Nat Biotechnol. (2016) and described further in Sanson, K.R., et al. Nat Commun (2018), which combine improved on-target activity predictions (Rule Set 2) with an off-target score, the Cutting Frequency Determination (CFD). The library is designed to be compact and efficient to maximize screening efficiency and performance.

Custom pools for follow-up screening or 10x Genomics Compatible CRISPR pools are also available by contacting your local sales representative.

Application

  • Functional Genomics/Target Validation
  • Unbiased wholed genome forward genetic screening
  • Validated positive and negative controls
  • Set up and optimization of screen assay

Features and Benefits

  • Focus on your research, and we will generate your lentivirus screening library
  • Use CRISPR nucleases to knockout protein-coding genes to assess their function.
  • Human Genes targeted 19,114
  • Compact library of ~ four gRNAs per gene (76,441 total)
  • Total Controls 1000(pools are gRNA-only, Cas9 sold separately) See products: LVCAS9BST or LVCAS9NEO for sources of Cas9

Principle

In a CRISPR KO screen, Cas9 introduces double-strand breaks at locations specified by a gRNA. When the endogenous non-homologous end-joining (NHEJ) DNA repair system corrects these breaks, this often leads to the introduction of frame-shift mutations that effectively knock out the gene. Thus, the power of CRISPR for genome engineering, coupled with the ability to perform large-scale, whole-genome loss-of-function (LOF) screening, has allowed breakthroughs in identifying gene pathways in drug resistance and disease.

Legal Information

CRISPR USE LICENSE AGREEMENT

LENTIVIRAL, WPRE AND EVROGEN FLUOROPHORE LICENSES

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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John G Doench et al.
Nature biotechnology, 34(2), 184-191 (2016-01-19)
CRISPR-Cas9-based genetic screens are a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target different sites in the genome with relative ease, but the on-target activity and off-target

Articles

The Researcher’s Guide to Designing CRISPR and RNAi Screening Experiments

This screening guide covers how to choose a cell line, a screening library, and experimental conditions as well as tips for designing and performing your experiment.

Genomic DNA Prep & Deconvolution Services

Deconvolute pooled shRNA samples post-screening to identify significant TRC clones with our service.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service