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PCRISPR006

Sigma-Aldrich

Human CRISPR Poison Exon Knockout Library

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About This Item

UNSPSC Code:
41105904
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, paired guide Poison (pgPoison) library targets 3` splice sites with paired guide RNAs for alternative exon removal (pgRNAs), induce skipping of "poison" cassette exons and corresponding upstream constitutive exons in ultraconserved regions. This compact library (targets 963 exons) allows direct comparison between loss of a constitutive coding exon (knockout) with loss of a poison exon.

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
  • Dual-gRNA
  • Poison exon
  • Internal controls included

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.
  • Built-in enrichment and depletion controls allow researchers to gauge the success of their pooled screening experiments confidently.
  • Dual-Guide Vector System (pools are gRNA-only, Cas9 sold separately) See products: LVCAS9BST or LVCAS9NEO.
  • Ease of optimization: Utilizes BFP and Puromycin as selection markers under EF1alpha promoter.

Principle

In traditional CRISPR KO screens, Cas9 introduces double-strand breaks at locations specified by a gRNA. In this library, a CRISPR-Cas9-based method is used by expressing two sgRNAs to manipulate isoforms independent of gene inactivation. This approach enables rapid suppression of exon recognition in polyclonal settings to identify functional roles for individual exons.

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

Certificates of Analysis (COA)

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James D Thomas et al.
Nature genetics, 52(1), 84-94 (2020-01-09)
While RNA-seq has enabled comprehensive quantification of alternative splicing, no correspondingly high-throughput assay exists for functionally interrogating individual isoforms. We describe pgFARM (paired guide RNAs for alternative exon removal), a CRISPR-Cas9-based method to manipulate isoforms independent of gene inactivation. This

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