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PCRISPR008

Sigma-Aldrich

CRISPR Toronto Knockout Library V3

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About This Item

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 Whole Genome Toronto KnockOut Library v3 (TKOv3) is optimized for editing efficiency using empirical data described in the findings by the Moffat lab.The compact library contains 70,948 gRNAs targeting 18,053 protein-coding genes (4 gRNAs/gene) with 142 control gRNAs targeting EGFP, LacZ, and luciferase (71,090 total). The optimized library size of TKOv3 offers improved accuracy, efficiency, and scalability for CRISPR KO screens.

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.
  • Resolve complex gene regulatory networks critical for biomarker or drug target discovery (pools are gRNA-only, Cas9 sold separately)
  • See products: LVCAS9BST or LVCAS9NEO for sources of Cas9.
  • Ease of optimization: Utilizes BFP and Puromycin as selection markers under EF1alpha promoter.

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 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Evaluation and Design of Genome-Wide CRISPR/SpCas9 Knockout Screens.
Hart, et al.
G3 (Bethesda, Md.), 7, 2719-2727 (2018)
Barbara Mair et al.
Cell reports, 27(2), 599-615 (2019-04-11)
Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are indispensable for hPSC

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