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CRISPRa and CRISPRi Human Genome-Wide Libraries

Engineered versions of deactivated Cas9 (dCas9) have proven to be effective modulators of gene expression when complexed with repressor (e.g., dCas9-KRAB) or activator (e.g., dCas9-VPH) proteins and targeted to the upstream promoter regions of gene targets with sgRNA. This technology forms the basis of the “CRISPR interference” (CRISPRi) and CRISPR activation (CRISPRa) systems. To enable CRISPRa or CRISPRi screens, Cellecta offers:

  • Pooled genome-wide lentiviral CRISPRa and CRISPRi libraries for screens targeting almost 19,000 human genes
  • Libraries which comprise two sgRNAs targeting each gene for stronger activation and repression (dual-sgRNA constructs library)
  • Both plasmid (200ug) and pre-packaged lentiviral particle (2x 10^8 TU & 1x 10^9 TU) formats available

 

CRISPRa targeted gene activation and CRISPRi targeted gene repression provide alternative genetic screening approaches to identify genes required to maintain a biological response or, with CRISPRa, genes whose activation initiates a response—a gain-of-function screen. Cellecta offers pre-made, pooled lentiviral libraries targeting all human protein-coding genes for both types of screens.

Dual sgRNA CRISPRa Library Activates More Genes

Two CRISPRa sgRNA to the Same Target Increase Expression. Two different sgRNA targeting the MyoD gene were transduced separately and together in triplicate into MDA231 cells.  One of the guides activated expression poorly by itself, and the other induced a bit over 10-fold activation. However, when the both guides were combined in the same cell, overall induction increased by almost an additional 10-fold.

Many gene activators and repressors complex with the promoter region of their target genes in multiple places. Cellecta has found that using this multiplex approach enhances the gene modulation effect, particularly with CRISPRa.

CRISPRa Single vs. Dual sgRNA to Same Gene

 

Increased Gene Activation with Dual-sgRNA. To detect whether the CRISPRa dual-sgRNA Library increases the levels of gene activation as compared to the single-sgRNA CRISPRa library, several hundred cells transduced with either the standard single sgRNA library or the dual-sgRNA library were sorted by FACS into small populations of ca. 10 cells/well. Both genomic DNA and RNA was isolated from each set of cells. The sgRNA library constructs in the cells were identified from the genomic DNA, and the DriverMap Targeted RNA-Sequencing Assay was used to assess the expression level of all human protein-coding genes. The data was then analyzed to correlate the expression of the targets for the sgRNAs identified in each cell group. The comparison showed that 33% of the gene targets were not induced with the single-sgRNA library, as compared with 27% with the dual-sgRNA CRISPRa Library, and, correspondingly, 7% more genes showed an induction of greater than 10-fold with the dual-sgRNA CRISPRa library.  

Cellecta’s CRISPRa and CRISPRi human genome-wide sgRNA libraries have been designed to target almost 19,000 human protein-coding genes using 5 sgRNAs designed against the promoter region based on Horbeck, et al. (eLife 2016;5:e19760 DOI: 10.7554). Our dual-guide CRISPRa and CRISPRi libraries use the same 5 effective sgRNA in dual combination on each of 5 constructs (sgRNA 1 & 2, sgRNA 1 & 3, sgRNA, sgRNA 2 & 3, etc.). Libraries are available in 200 ug plasmid and pre-packaged 2 x 10^8 TU and 1 x 10^9 TU lentiviral particles.

Human Genome-Wide CRISPRa Single-sgRNA and Dual-sgRNA Libraries

Selective activation of targeted genes when used in cells expressing dCas9-VPH (or other dCas9-activator variations compatible with spCas9 sgRNA designs)

Human Genome-Wide CRISPRi Single-sgRNA and Dual-sgRNA Libraries

Selective expression inhibition of targeted genes when used in cells expressing dCas9-KRAB (or other deactivated Cas9 repressors compatible with spCas9 sgRNA designs)

Mouse Genome-Wide CRISPRa and CRISPRi libraries are also available.

 Ordering information

 

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

Quantity provided in each library: 200 ug plasmid; 2 x 10^8 TU or 1 x 10^9 TU packaged

Human Genome-Wide CRISPRa sgRNA Library (single-guide)

Catalog#NameQuantityPriceBuy
KAHGW-106-PHuman Genome-Wide CRISPRa sgRNA Library (plasmid)200 ug$10,000.00
KAHGW-106-V8Human Genome-Wide CRISPRa sgRNA Library (virus)2 x 10^8 TU$8,500.00
KAHGW-106-V9Human Genome-Wide CRISPRa sgRNA Library (virus)1 x 10^9 TU$18,000.00

Human Genome-Wide CRISPRa Dual-sgRNA Library (dual-guide)

Catalog#NameQuantityPriceBuy
KADHGW-105K-PHuman Genome-Wide CRISPRa Dual-sgRNA Library (plasmid)200 ug$12,000.00
KADHGW-105K-V8Human Genome-Wide CRISPRa Dual-sgRNA Library (virus)2 x 10^8 TU$9,500.00
KADHGW-105K-V9Human Genome-Wide CRISPRa Dual-sgRNA Library (virus)1 x 10^9 TU$20,000.00

Human Genome-Wide CRISPRi sgRNA Library (single-guide)

Catalog#NameQuantityPriceBuy
KIHGW-106-PHuman Genome-Wide CRISPRi sgRNA Library (plasmid)200 ug$10,000.00
KIHGW-106-V8Human Genome-Wide CRISPRi sgRNA Library (virus)2 x 10^8 TU$8,500.00
KIHGW-106-V9Human Genome-Wide CRISPRi sgRNA Library (virus)1 x 10^9 TU$18,000.00

Human Genome-Wide CRISPRi Dual-sgRNA Library (dual-guide)

Catalog#NameQuantityPriceBuy
KIDHGW-105K-PHuman Genome-Wide CRISPRi Dual-sgRNA Library (plasmid)200 ug$12,000.00
KIDHGW-105K-V8Human Genome-Wide CRISPRi Dual-sgRNA Library (virus)2 x 10^8 TU$9,500.00
KIDHGW-105K-V9Human Genome-Wide CRISPRi Dual-sgRNA Library (virus)1 x 10^9 TU$20,000.00

If you are interested in a custom library, Cellecta can help. Visit our Order page for specific instructions.

Flyer

User Manual

Gene Lists

sgRNA Sequences

  • Sequences are provided upon shipment of library.

Vector Information

Single sgRNA Library VectorNGS CassetteSequence
(FASTA|GenBank|SnapGene*)
Map
pRSG16-U6-sg-HTS6C-UbiC-TagRFP-2A-PuroHTS6C.txt | .gb | .dnaMap
Dual-sgRNA Library VectorNGS CassetteSequence
(FASTA|GenBank|SnapGene*)
Map
pRSL10-mU6-sgRNA1-hU6-sgRNA2-UbiC-TagRFP-2A-PuropRSL10.txt | .gb | .dnaMap

Product Certificates

Lot NumberCatalog NumberDescriptionQuantityProduct Certificate
190724001KADHGW-105K-PHuman Genome-Wide CRISPRa Dual-sgRNA Library (plasmid)200 µgPDF
190417035KIHGW-106-V8Human Genome-Wide CRISPRi sgRNA Library (virus)2 x 108 TUEmail us
190327001KAHGW-106-V9Human Genome-Wide CRISPRa sgRNA Library (virus)1 x 109 TUPDF
181128001KIHGW-106-V8Human Genome-Wide CRISPRi sgRNA Library (virus)2 x 108 TUPDF
180124001KIHGW-106-PHuman Genome-Wide CRISPRi sgRNA Library (plasmid)200 µgPDF
18011730KAHGW-106-V8Human Genome-Wide CRISPRa sgRNA Library (virus)2 x 108 TUPDF
18011702KIHGW-106-V8Human Genome-Wide CRISPRi sgRNA Library (virus)2 x 108 TUPDF
17122701KAHGW-106-PHuman Genome-Wide CRISPRa sgRNA Library (plasmid)200 µgPDF
 

Safety Data Sheets (SDS)

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.

dCas9 Expression Plasmids

More information >

Catalog#NameQuantityPriceBuy
SVKRABC9B-PSCRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (plasmid)25 ug$300.00
SVKRABC9E2B-PSCRISPRi dCas9-KRAB pRDKCE2B-EFS-dCas9-KRAB-2A-Blast (plasmid)25 ug$300.00
SVKRABC9H-PSCRISPRi dCas9-KRAB pRDKCCH-CMV-dCas9-KRAB-2A-Hygro (plasmid)25 ug$300.00
SVKRABC9RB-PSCRISPRi dCas9-KRAB pRDKCRB-RSV-dCas9-KRAB-2A-Blast (plasmid)25 ug$300.00
SVKRABC9SFB-PSCRISPRi dCas9-KRAB pRDKCSFB-SFFV-dCas9-KRAB-2A-Blast (plasmid)25 ug$300.00
SVKRABC9B-VSCRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (virus)1 x 10^6 TU$500.00
SVKRABC9E2B-VSCRISPRi dCas9-KRAB pRDKCE2B-EFS-dCas9-KRAB-2A-Blast (virus)1 x 10^6 TU$500.00
SVKRABC9H-VSCRISPRi dCas9-KRAB pRDKCCH-CMV-dCas9-KRAB-2A-Hygro (virus)1 x 10^6 TU$500.00
SVKRABC9RB-VSCRISPRi dCas9-KRAB pRDKCRB-RSV-dCas9-KRAB-2A-Blast (virus)1 x 10^6 TU$500.00
SVKRABC9SFB-VSCRISPRi dCas9-KRAB pRDKCSFB-SFFV-dCas9-KRAB-2A-Blast (virus)1 x 10^6 TU$500.00
Catalog#NameQuantityPriceBuy
SVVPHC9B-PSCRISPRa dCas9-VPH pRDVCCB-CMV-dCas9-VPH-2A-Blast (plasmid)25 ug$300.00
SVVPHC9E2B-PSCRISPRa dCas9-VPH pRDVCE2B-EFS-dCas9-VPH-2A-Blast (plasmid)25 ug$300.00
SVVPHC9RB-PSCRISPRa dCas9-VPH pRDVCRB-RSV-dCas9-VPH-2A-Blast (plasmid)25 ug$300.00
SVVPHC9SFB-PSCRISPRa dCas9-VPH pRDVCSFB-SFFV-dCas9-VPH-2A-Blast (plasmid)25 ug$300.00
SVVPHC9B-VSCRISPRa dCas9-VPH pRDVCCB-CMV-dCas9-VPH-2A-Blast (virus)1 x 10^6 TU$500.00
SVVPHC9E2B-VSCRISPRa dCas9-VPH pRDVCE2B-EFS-dCas9-VPH-2A-Blast (virus)1 x 10^6 TU$500.00
SVVPHC9RB-VSCRISPRa dCas9-VPH pRDVCRB-RSV-dCas9-VPH-2A-Blast (virus)1 x 10^6 TU$500.00
SVVPHC9SFB-VSCRISPRa dCas9-VPH pRDVCSFB-SFFV-dCas9-VPH-2A-Blast (virus)1 x 10^6 TU$500.00

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

Catalog#NameQuantityPriceBuy
CPCP-K2AReady-to-Use Lentiviral Packaging Plasmid Mix250 ug$340.00
LNGS-120NGS Prep Kit for sgRNA Libraries in pRSG16/17 (KOHGW) with Alignment SoftwareKit for 6-48 samples (48 preps of 50ug each, 12 multiplex)$950.00
LNGS-120-SPSupplementary Primer Set for LNGS-120 (12 Additional Index Primers)48 Reactions (12 Index Primers x 4 reactions each)$475.00
LFVC1LentiFuge™ Viral Concentration Reagent1 ml of 1000x$325.00
LTDR1LentiTrans™ Transduction Reagent (Polybrene for Transduction)1 ml of 1000x$175.00
LTSET-GLentiPrep™ Lentiviral Reagent Set with GFP Control for Packaging and TransductionKit$750.00
LTSET-RLentiPrep™ Lentiviral Reagent Set with RFP Control for Packaging and TransductionKit$750.00
CRATESTCRISPRaTest™ Functional dCas9-Activator Assay Kit1 kit (for 5-15 cell lines)$850.00
CRITESTCRISPRiTest™ Functional dCas9-Repressor Assay Kit1 kit (for 5-10 cell lines)$850.00
PROMTESTPromoterTest™ Assay Kit (8 promoter constructs)1 kit (for 10 cell lines)$895.00