Pooled Libraries
Dual sgRNA CRISPRi library

Dual sgRNA CRISPRi Libraries
(Pooled Library #187246, #187247, #187248, #1000000192)

Purpose

Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette.
Vector Backbone

pJR104

Depositing Labs

Jonathan Weissman
Publication

Replogle et al bioRxiv 2022.07.13.499814. doi: 10.1101/2022.07.13.499814. (How to cite )

Ordering

Item	Catalog #	Description	Quantity
Pooled Library	1000000192	Three tubes, each with a third of the library (Library 1-2, 3-4, and 5-6) in backbone pBA904 #187246 (Library 1-2) #187247 (Library 3-4) #187248 (Library 5-6)	1	$1200	Add to Cart
Pooled Library	187246	hCRISPRi_dual_1_2 pooled library^†	1	$500	Add to Cart
Pooled Library	187247	hCRISPRi_dual_3_4 pooled library^†	1	$500	Add to Cart
Pooled Library	187248	hCRISPRi_dual_5_6 pooled library^†	1	$500	Add to Cart

Available to Academic and Nonprofits Only

^† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pHR-UCOE-SFFV-dCas9-KRAB(Kox1)-MeCP2-P2A-EGFP (Addgene #188902) or Zim3-dCas9.

Library Details

Genes/gRNAs/plasmids
22,580 dual sgRNA elements
Controls
1,026 dual sgRNA control elements
Lentiviral Generation
3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.

Volume
∼20 µL
Concentration
50 ng/µL

Resource Information

Protocols
- Illumina Sequencing Sample Preparation (369 KB)
- Liquid Culture Re-Amplification Protocol (71 KB)
- Cell Line Creation and Validation Primer (148 KB)
- CRISPRi/a Custom Library Cloning Protocol (406 KB)
- For more links and protocols, please see the Weismann lab's CRISPRi/a screening materials page.
Depositor Data
- List of CRISPRi sgRNAs (7.4 MB)
Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials

Depositor Comments

User instructions are available on GitHub ( https://github.com/josephreplogle/CRISPRi-dual-sgRNA-screens).

How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

For your Materials & Methods section:

Dual sgRNA CRISPRi libraries were a gift from Jonathan Weissman (Addgene #)
For your References section:

Maximizing CRISPRi efficacy and accessibility with dual-sgRNA libraries and optimal effectors. Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M. bioRxiv 2022.07.13.499814. doi: 10.1101/2022.07.13.499814. bioRxiv

Dual sgRNA CRISPRi Libraries (Pooled Library #187246, #187247, #187248, #1000000192)