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PCRISPR003

Sigma-Aldrich

Human CRISPR Inhibition Dolcetto Library

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packaging

pkg of 10 vials (5x200µL aliquots of each component)

concentration

≥5x108 VP/ml (via p24 Assay)

application(s)

CRISPR

shipped in

dry ice

storage temp.

−70°C

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PCRISPR007PCRISPR001PCRISPR002
concentration

≥5x108 VP/ml (via p24 Assay)

concentration

≥5x108 VP/ml (via p24 Assay)

concentration

≥5x108 VP/ml (via p24 Assay)

concentration

≥5x108 VP/ml (via p24 Assay)

application(s)

CRISPR

application(s)

CRISPR

application(s)

CRISPR

application(s)

CRISPR

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

storage temp.

−70°C

storage temp.

−70°C

storage temp.

−70°C

storage temp.

−70°C

General description

The Human CRISPR inhibition (CRISPRi) library (Dolcetto) is a single compact library that inhibits over 18,000 human genes and is used for genome-wide inhibition screening using a KRAB-dCas9 effector. The library is designed to be compact and efficient to maximize screening efficiency and performance, as published by Sanson, K.R., et al. Nat Commun 9, 5416 (2018).

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
  • Focused forward genetic activation screening
  • Validated positive and negative controls
  • Set up and optimization of screen assay
  • Creation of cell lines stably expressing KRAB-dCas9

Features and Benefits

  • Focus on your research, and we will generate your lentivirus screening library
  • Compact and ready to use CRISPRi library ~6 sgRNAs
  • Optimized (F+E) gRNA scaffold (pools are gRNA-only, KRAB-dCas9 sold separately)
  • Can be paired with CRISPRICON to ease optimization using validated positive control targeting RAB1A and non-targeting control for assay setup
  • UCOE KRAB-dCas9 for consistent effector expression across a wide variety of cell lines
  • Ease of optimization: Utilizes BFP and Puromycin as selection markers under EF1alpha promoter.

Components

2 Subpools with a minimum concentration of 5x108 VP/mL(via p24 assay)
PCRISPR003A - Human CRISPR Inhibition Dolcetto Library Set A
PCRISPR003B - Human CRISPR Inhibition Dolcetto Library Set B

Principle

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. CRISPR is most commonly used to create double-stranded breaks that often result in loss of gene function (CRISPR-KO). However, the full extent of CRISPR′s utility extends beyond just targeted cutting of DNA. CRISPR′s Nuclease-independent applications provide equal targeting specificity. Instead of cutting, CRISPRi allows for targeted interference of gene function by delivering transcriptional repressor domains to a specific target sequence using modified dCas9 + gRNA complexes. Gene knockdown complements CRISPR-KO and CRISPRa (activation) and has distinct advantages over existing loss-of-function strategies like RNAi.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Kendall R Sanson et al.
Nature communications, 9(1), 5416-5416 (2018-12-24)
The creation of genome-wide libraries for CRISPR knockout (CRISPRko), interference (CRISPRi), and activation (CRISPRa) has enabled the systematic interrogation of gene function. Here, we show that our recently-described CRISPRko library (Brunello) is more effective than previously published libraries at distinguishing

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