Brunello Whole Genome KO CRISPR Pooled Lentiviral Library (1)
- 1 vector system: contains Cas9 in addition to gRNA
- ~ 5 gRNAs/gene for editing 19,114 genes.
Calabrese Whole Genome Activator CRISPR Pooled Lentiviral Library (2)
- The human CRISPR activation library (Calabrese P65-HSF) activates over 18,000 human genes and is used for genome-wide activation screening.
- Each activated gene is targeted by 3-6 gRNAs.
Dolcetto Whole Genome Inhibitor CRISPR Pooled Lentiviral Library (2)
- The human CRISPR inhibition library (Dolcetto) is in the backbone pXPR_050 (VP64) and inhibits over 18,000 human genes.
- Each inhibited gene is targeted by 3-6 gRNAs.
Services provided include library generation, killing curve with puromycin, CRISPR screen in the absence or presence small molecules, optimization of pcrs to quantify distribution of sgRNA before/after selection.
Coordination with GCCRI NGS (Zhao Lai) and Bioinformatic (YiDong Chen) Facilities for sequencing and data analysis.
1. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9 . Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. 10.1038/nbt.3437
2. Optimized libraries for CRISPR-Cas9 genetic screens with multiple modalities. Sanson KR, Hanna RE, Hegde M, Donovan KF, Strand C, Sullender ME, Vaimberg EW, Goodale A, Root DE, Piccioni F, Doench JG. Nat Commun. 2018 Dec 21;9(1):5416.