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SRX1180259: Optimized sgRNA design to maximize activity and minimize off-target effects for genetic screens with CRISPR-Cas9
3 ILLUMINA (Illumina HiSeq 2500) runs: 367.8M spots, 18.4G bases, 7.8Gb downloads

Submitted by: (mhegde@broadinstitute.org)
Study: Optimized sgRNA design to maximize activity and minimize off-target effects for genetic screens with CRISPR-Cas9
show Abstracthide Abstract
CRISPR technology has proven to be a powerful means of conducting genetic screens. The ease of programming the Cas9 protein with an sgRNA presents an abundance of potential target sites for each gene, but the on-target activity and off-target effects of individual sgRNAs can vary. Here, we incorporated recently devised sgRNA design rules to create human and mouse genome-wide libraries, performed positive and negative selection screens, and observed that the use of these rules greatly enhances the consistency and depth of screening results. Additionally, we profiled the off-target activity of thousands of sgRNAs and developed a metric to predict off-target sites that outperforms existing metrics. We combine these findings from large-scale, empirical data to improve our computational design rules and create highly-optimized sgRNA libraries that maximize on-target activity and minimize off-target effects in order to enable more effective and efficient genetic screens and genome engineering.
Sample:
SAMN04027369 • SRS1053777 • All experiments • All runs
Organism: Homo sapiens
Library:
Name: HT29 A375
Instrument: Illumina HiSeq 2500
Strategy: AMPLICON
Source: SYNTHETIC
Selection: PCR
Layout: SINGLE
Spot descriptor:
forward

Runs: 3 runs, 367.8M spots, 18.4G bases, 7.8Gb
Run# of Spots# of BasesSizePublished
SRR224328611,703,014585.2M254.4Mb2015-09-10
SRR2299054187,316,3379.4G3.9Gb2015-09-19
SRR2431464168,810,8198.4G3.6Gb2015-09-18

ID:
1712129

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