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)
NOTE: Cellecta offers academic discounts on many of our pre-made libraries to qualifying academic institutions. Please email email@example.com using your institutional email account, and ask for a quote with the academic discount pricing.
We can also prepare custom libraries according to your specifications. Email firstname.lastname@example.org for details or to obtain a quote.
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.
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.
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.
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.
|CRISPR Library Vector||NGS Cassette||Sequence|
|pRSG16-U6-sg-HTS6C-UbiC-TagRFP-2A-Puro||HTS6C||.txt | .gb | .dna||Map|
|pRSGEP-U6-sg-HTS6C-EF1-Puro||HTS6C||.txt | .gb | .dna||Map|
Lot#-Specific Product Information (Product Analysis Certificates/PACs)
- CRISPR Human Genome 80K Knockout Library in pRSG16:
Safety Data Sheet (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 barcodes, sequence, 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.
|Catalog #||Item Description||Price|
|CPCP-K2A||Ready-to-Use Lentiviral Packaging Plasmid Mix, 250 μg|
(for (25) 10-cm plates)
|LFVC1||LentiFuge™ Viral Concentration Reagent, 1 ml|
(for 1 liter supernatant)
|CRUTEST||CRISPRuTest™ Functional Cas9 Activity Assay Kit (Mammalian)||$850|
|LNGS-120||NGS Prep Kit for KOHGW CRISPR sgRNA libraries||$875|