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Single-Module Human Genome-Wide CRISPR Library (80K)

Cellecta’s Single-module Human Genome-Wide CRISPR Library was designed to be small enough (~80,000 constructs) to enable convenient genome-wide “dropout viability” screens with most standard human cell modules. With 4 sgRNAs targeting each gene, all of which have been designed using the latest criteria for effective guides described by Doench, et al. and containing the Cellecta-designed HEAT sgRNA structure which we have proven to generate more robust screening results than the standard sgRNA design, the library has deep enough gene coverage to generate efficient results.

Genome-Wide Coverage with a Convenient Single 80K Module

  • Single library with ~80,000 constructs targeting approximately 19,000 protein-coding genes
  • Each gene is targeted by 4 sgRNAs designed based on Doench, et al. criteria
  • Targets all human protein-coding genes—over 19,000 gene targets.

The 80K Single-Module Human Genome-Wide CRISPR Library includes the following controls

  • standard non-targeting controls
  • non-specific control (sgRNA targeting introns throughout the genome)
  • lethal sgRNA targeting generally essential genes (i.e., positive controls for dropout screens)

Ordering information

NOTE: Cellecta offers academic discounts on many of our plasmid and pre-packaged libraries to qualifying academic institutions. Please email us at orders@cellecta.com using your institutional email account, and ask for a quote with the discounted pricing.

Catalog#NameCategoryPriceBuy
KOHGW-80K-PCRISPR Human Genome 80K Knockout Library (plasmid)$10,000.00
KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)$8,500.00
KOHGW-80K-V9CRISPR Human Genome 80K Knockout Library (virus)$18,000.00
KOHGW-EP-80K-PCRISPR Human Genome 80K Knockout Library in EF1-Puro Vector (plasmid)$10,000.00
KOHGW-EP-80K-V8CRISPR Human Genome 80K Knockout Library in EF1-Puro Vector (virus)$8,500.00
KOHGW-EP-80K-V9CRISPR Human Genome 80K Knockout Library in EF1-Puro Vector (virus)$18,000.00

We can also prepare custom libraries according to your specifications. Email sales@cellecta.com for details or to obtain a quote.

Single-module Human Genome-Wide CRISPR Library

To identify the best performing sgRNA design, we built an sgRNA design optimization library of different handle structures, all targeting the same ~900 human genomic sites. Targeted sites included essential genes, non-essential genes, introns, and non-human control sequences. In order to identify the best performing sgRNA design, the optimization library was transduced into CAS9 expressing cell lines validated with CRISPRTest Cas9 Assay Kit. Samples were then collected at multiple time points post-transduction. The waterfall plots display the depletion fold of positive and negative controls within the library for each design.

 

Z-scores of sgRNA handle designs

Based on the calculated depletion folds of positive controls in the design optimization screen (Fig. 1), Z-scores were calculated for each sgRNA handle design. Following analysis of the depletion values and Z-scores across all the tested cell lines at multiple time points, the HEAT design was identified as the best performing and selected for the construction of Cellecta’s pre-made and custom CRISPR sgRNA libraries.

 

Library Representation - Human 80K

The histogram above shows the distribution of sgRNA sequences in the CRISPR Human Genome 80K Knockout Library with RFP-Puro (Cat.# KOHGW-80K-P). Virtually all of the sgRNAs are present in more than 500 copies and most are around 2,000 copies per 100 million NGS reads. Also, there is a 3.2-fold difference between the most and least represented sequences for 90% of the sgRNAs, an indication that the library has a balanced representation of all sgRNAs.

 

Single-module Human Genome-Wide CRISPR Library Kit image

Our CRISPR Human Genome 80K Knockout Library (plasmid) is provided in a single tube of plasmid library DNA, ready to package. Cellecta also provides pre-packaged libraries.

 

User Manual

Gene List

sgRNA Sequences

  • Sequences are provided upon shipment of library.

Vector Information

CRISPR Library VectorNGS CassetteSequence
(FASTA|GenBank|SnapGene*)
Map
pRSG16-U6-sg-HTS6C-UbiC-TagRFP-2A-PuroHTS6C.txt | .gb | .dnaMap
pRSGEP-U6-sg-HTS6C-EF1-PuroHTS6C.txt | .gb | .dnaMap

Product Certificates

Lot NumberCatalog NumberDescriptionQuantityProduct Certificate
180731016KOHGW-EP-80K-V9CRISPR Human Genome 80K Knockout Library in EF1-Puro Vector (virus)1 x 109 TUPDF
180228049KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF
17122020KOHGW-EP-80K-PCRISPR Human Genome 80K Knockout Library in EF1-Puro Vector (plasmid)200 µgPDF
17050302KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF
17050301KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF
17031537KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF
17022305KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF
16110923KOHGW-80K-PCRISPR Human Genome 80K Knockout Library (plasmid)200 µgPDF
16110922KOHGW-80K-V8CRISPR Human Genome 80K Knockout Library (virus)2 x 108 TUPDF

Safety Data Sheets (SDS)

* Download the SnapGene® Viewer to open .dna format files.

Next-Gen Sequencing and Analysis

Cellecta CRISPR sgRNA Libraries are provided with a complete protocol and all sequencing information to enable researchers to perform experiments and extract the data. However, we also provide Next-Gen Sequencing and analysis services for researchers running their own screens with our libraries. Just provide harvested cells or tissue for each time point or treatment condition (one sample), and we’ll extract DNA, amplify the sgRNA sequences, perform NGS, and assemble the data with some basic analysis.

You provide Cellecta with frozen cell pellets or tissue after screening…

…and Cellecta does the rest:

  • Extracts genomic DNA
  • Amplifies sgRNA
  • Performs NGS on the Illumina NextSeq or HiSeq
  • Enumerates sgRNA counts from raw sequencing data

Please see the Next-Gen Sequencing and Analysis web page for additional information and instructions on how to order.

Cas9 Expression Plasmids

More information >

Catalog#NameCategoryPriceBuy
SVC9-PSCas9 Plasmids (targeted gene knockouts and screening using CRISPR) - Plasmid$300.00
SVC9-VSCas9 Plasmids (targeted gene knockouts and screening using CRISPR) - Virus$500.00
SVCOPG-PCRISPR Target for sgCopGFP Control (pRCdsGEB-CMV-dsCopGFP-EF1-Bleo, plasmid)$700.00
SVCOPG-VCRISPR Target for sgCopGFP Control (pRCdsGEB-CMV-dsCopGFP-EF1-Bleo, virus)$500.00

dCas9 Expression Plasmids

More information >

Catalog#NameCategoryPriceBuy
SVKRABC9-PS-plsamidCRISPRa dCas9-KRAB (For use with the CRISPR gene inhibition) - plasmid$300.00
SVKRABC9B-VS-virusCRISPRa dCas9-KRAB (For use with the CRISPR gene inhibition) - virus$500.00

Catalog#NameCategoryPriceBuy
SVVPHC9B-PS-plasCRISPRa dCas9-VPH (For use with the CRISPR gene activation) - plasmid$300.00
SVVPHC9B-VS-virusCRISPRa dCas9-VPH (For use with the CRISPR gene activation) - virus$500.00

We can also provide other configurations of Cas9 or dCas9 expression plasmids on a custom basis. Please inquire if you require something different than the items described above.

Catalog #Item DescriptionPrice
CPCP-K2AReady-to-Use Lentiviral Packaging Plasmid Mix, 250 μg
(for (25) 10-cm plates)
$325
LFVC1LentiFuge™ Viral Concentration Reagent, 1 ml
(for 1 liter supernatant)
$325
CRUTESTCRISPRuTest™ Functional Cas9 Activity Assay Kit (Mammalian)$850
LNGS-120NGS Prep Kit for KOHGW CRISPR sgRNA libraries $875