CHyMErA Knockout Libraries
(Pooled Library #155199, #155200)
The Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA) lentiviral libraries express hybrid Cas9-Cas12a guide RNAs under a single U6 promoter.
The Paralog & Dual-targeting hybrid guide RNA Library targets human paralog pairs with single- and combinatorial-targeting hgRNAs and can be used to identify genetic interactions between paralogs. The library also contains single- and dual-targeting hybrid guides that target ~5,000 highly expressed genes permitting capture of gene ablation phenotypes with increased sensitivity.
The Exon-deletion hgRNA Library deletes human alternative cassette exons and disrupts genes containing alternative exons, allowing investigation into alternative exon function.
pLCHKO - does not express Cas9 or Cas12a
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||155199||CHyMErA Paralog & Dual-targeting hgRNA pooled library†||1||$540||Add to Cart|
|Pooled Library||155200||CHyMErA Exon-deletion hgRNA pooled library†||1||$540||Add to Cart|
† SpCas9 and LbCas12a plasmids are NOT included with this item and will have to be ordered separately. Can be used in conjunction with lenti-Cas9-2A-BLAST (Addgene #73310) and plenti-Lb-Cas12a-2xNLS (Addgene #155046), or with a cell line expressing both SpCas9 and LbCas12a.
Paralog & Dual-targeting library: 6,333 genes (1,344 paralogs & 4,993 genes)
Exon-deletion library: 2,157 alternative cassette exons in 1,745 genes
Paralog & Dual-targeting library: 92,746
Exon-deletion library: 91,948
Paralog & Dual-targeting library: 4,993 intergenic and non-targeting hgRNAs
Exon-deletion library: 1,647 intergenic control hgRNAs
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.
Terms and Licenses
Screening analysis is described in:
Pooled Lentiviral CRISPR-Cas9 Screens for Functional Genomics in Mammalian Cells. Aregger M, Chandrashekhar M, Tong AHY, Chan K, Moffat J Methods Mol Biol. 2019;1869:169-188. doi: 10.1007/978-1-4939-8805-1_15. PMID 30324523
The depositing lab has provided analysis algorithms and R code in their Github repositories:
CHyMERa Dual-targeting Scoring
CHyMERa Exon-deletion Scoring
Read depositor Jason Moffat's description of CHyMErA in this Nature Bioengineering Behind The Paper article.
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:CHyMErA Knockout library was a gift from Jason Moffat and Benjamin Blencowe (Addgene #)
For your References section:Genetic interaction mapping and exon-resolution functional genomics with a hybrid Cas9-Cas12a platform. Gonatopoulos-Pournatzis T, Aregger M, Brown KR, Farhangmehr S, Braunschweig U, Ward HN, Ha KCH, Weiss A, Billmann M, Durbic T, Myers CL, Blencowe BJ, Moffat J. Nat Biotechnol. 2020 May;38(5):638-648. doi: 10.1038/s41587-020-0437-z. Epub 2020 Mar 16. PubMed 32249828