Three-Module Human Genome-Wide CRISPR Library
Cellecta’s original Genome-Wide CRISPR Knockout Pooled sgRNA Library was designed as three modules, each targeting approximately a third of the genes with eight sgRNAs per gene, which is more than most other libraries use. In addition to filtering to ensure guides are unique for each gene target to minimize false positives, the sgRNAs in Cellecta’s Three-Module Human Genome-Wide CRISPR Library incorporate Cellecta’s HEAT design sgRNA structure which has been shown to generate more robust screening results than the standard sgRNA design.
Deep Genome-Wide Coverage of All Human Genes
- 3 Modules, each covering approximately 6,300 genes
- Each gene is targeted by up to 8 sgRNA for a total of approximately 50,000 sgRNA per module
- Targets all human protein-coding genes—over 19,000 gene targets.
Three separate modules, each with approximately fifty thousand constructs provide a practical approach to screens with more guides per target. The modular format is similar to Cellecta’s Human Genome-Wide shRNA Library, and each of the CRISPR and RNAi Library modules target the same sets of genes so that investigators can more easily make use of both techniques for genome-wide loss of function screens.
Depending on the needs of the experiment, each of the three CRISPR Library modules can be used separately or combined for a single genome-wide knockout screen. For example, screens with a strong selection and few surviving cells may be easily run using a single large pool with all three modules combined. However, with “dropout viability” screens that require maintenance of hundreds of millions of cells for weeks at a time, individual screens with each module run separately, may be more practical.
The selection of genes targeted with each module also allows researchers primarily interested in therapeutic applications to focus on the most relevant and annotated set of disease related targets by screen Modules 1 and/or 2. The complete list of genes targeted by each modules is available below.
- CRISPR Human Genome Knockout Module 1 targets the most annotated genes, including members of major signal transduction pathways, genes from the Cancer Genome Atlas (TCGA), and approved drug targets.
- Module 2 targets additional disease-associated genes and drug targets
- Module 3 targets cell surface markers, extracellular matrix genes, and DNA binding genes
The target gene lists for the Three-Module Human Genome-Wide CRISPR Library can be downloaded here:
- CRISPR Human Genome KO Library Module 1
- CRISPR Human Genome KO Library Module 2
- CRISPR Human Genome KO Library Module 3
|CRISPR Library Vector||NGS Cassette||Sequence (.txt | .gbk | .dna*)||Map|
|pRSG16-U6-sg-HTS6C-UbiC-TagRFP-2A-Puro||HTS6C||FASTA | GenBank | SnapGene||Map|
* Download the SnapGene® Viewer to open .dna format files.
Next-Gen Sequencing and Analysis
Cellecta libraries are provided with a complete protocol and all sequencing information to enable researchers to perform high throughput genetics screens and analysis. However, we also do provide Next-Gen Sequencing and analysis services for researchers running their own screens with our libraries. Just provide harvested cells for each time point or treatment condition (one sample), and we extract DNA, amplify, 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
- Enumerate sgRNA counts from raw sequencing data
- Additional bioinformatics and pathway analysis available
Please see the Next-Gen Sequencing and Analysis web page for additional information and how to order.
|Catalog #||Item Description||Amount Provided|
|KOHGW-M1-P||CRISPR Human Genome KO Library, Module 1 (plasmid)||200 μg|
|KOHGW-M2-P||CRISPR Human Genome KO Library, Module 2 (plasmid)||200 μg|
|KOHGW-M3-P||CRISPR Human Genome KO Library, Module 3 (plasmid)||200 μg|
Packaged Viral Library:
|Catalog #||Item Description||Amount Provided|
|KOHGW-M1-V8||CRISPR Human Genome KO Library, Module 1 (packaged)||2 x 108 TU|
|KOHGW-M2-V8||CRISPR Human Genome KO Library, Module 2 (packaged)||2 x 108 TU|
|KOHGW-M3-V8||CRISPR Human Genome KO Library, Module 3 (packaged)||2 x 108 TU|
|KOHGW-M1-V9||CRISPR Human Genome KO Library, Module 1 (packaged)||1 x 109 TU|
|KOHGW-M2-V9||CRISPR Human Genome KO Library, Module 2 (packaged)||1 x 109 TU|
|KOHGW-M3-V9||CRISPR Human Genome KO Library, Module 3 (packaged)||1 x 109 TU|
What you will receive in addition to your order:
- Plasmid orders include 10 μg empty library vector, for cloning individual constructs used to validate hits from your screen.
- Virus orders include an additional small aliquot of virus to be used for titering in your target cells.
Please email email@example.com for questions or to obtain a quote.
|Catalog #||Item Description||Price|
|SVC9-PS||pR-CMV-Cas9-2A-Hygro Cas9 Expression Vector (plasmid), 25 μg||$300|
|SVC9-VS||pR-CMV-Cas9-2A-Hygro Cas9 Expression Vector (virus), 1 x 106 TU||$500|
|SVC9B-PS||pR-CMV-Cas9-2A-Blast Cas9 Expression Vector (plasmid), 25 μg||$300|
|SVC9B-VS||pR-CMV-Cas9-2A-Blast Cas9 Expression Vector (virus), 1 x 106 TU||$500|
|SVC9G-PS||pR-CMV-Cas9-2A-GFP Cas9 Expression Vector (plasmid), 25 μg||$300|
|SVC9G-VS||pR-CMV-Cas9-2A-GFP Cas9 Expression Vector (virus), 1 x 106 TU||$500|
|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)
|CRTEST||CRISPRtest™ Functional Cas9 Activity Kit (Human)|
(for 5 cell line pairs)