BARBEKO sgRNA Library
(Pooled Library #174163)
BARBEKO, short for iBARed cytosine Base Editing-mediated gene KnockOut, leverages CRISPR cytosine base editors for genome-scale knockout screens in human cells by perturbing gene start codons, splice sites, or by introducing premature termination codons. It is integrated with iBAR, a design in the sgRNA scaffold for improving screening quality and efficiency.
Each sgRNA construct contains one iBAR sequence and each sgRNA is followed by four iBARs (214,008 sgRNAiBARs in total).
pLenti-sgRNA-Lib - does not express Cas9
This library is designed for gene knockout screens by using CRISPR cytosine base editors (CBE), but not Cas9. Library was re-constructed into lentiviral sgRNA-iBAR backbone according to the method in Zhu et al., 2019 PMID: 30678704.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||174163||BARBEKO sgRNA library†||1||$540||Add to Cart|
Non-targeting controls: 500
Negative controls targeting safe-harbor regions: 499
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.
The CBE-expressing cassette from pCMV_AncBE4max (Addgene #112094) was transferrred into a lentiviral backbone to create pLenti-AncBE4max (Addgene #174164). To efficiently package the CBE into lentivirus, concentrate the lentiviral supernatants before use.
For cell fitness screens, ZFCiBAR algorithm can be used and obtained from https://github.com/wolfsonliu/zfc. For positive selection screens, MAGeCKiBAR algorithm can be used for analysis (Zhu et al., 2019): https://bitbucket.org/WeiLab/mageck-ibar/.
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:BARBEKO sgRNA library was a gift from Wensheng Wei (Addgene #174163)
For your References section:Genome-wide interrogation of gene functions through base editor screens empowered by barcoded sgRNAs. Xu P, Liu Z, Liu Y, Ma H, Xu Y, Bao Y, Zhu S, Cao Z, Wu Z, Zhou Z, Wei W. Nat Biotechnol. 2021 Jun 21. pii: 10.1038/s41587-021-00944-1. doi: 10.1038/s41587-021-00944-1. PubMed 34155407