Integration profiling: a genome-wide mapping of sequence function

Tuesday, October 25, 2011 — Poster Session II
Noon – 2:00 p.m.	Natcher Conference Center	NICHD	GEN/GENOM-6

Authors

• Y Guo
• J Park
• E Humes
• K Hoe
• H Levin

Abstract

The DNA transposon Hermes, derived from the housefly, is highly active in Schizosaccharomyces pombe and disrupts ORFs. Hermes can thus be used as a mutagen in genetic screens. Using Hermes, we have developed a genome-wide method called integration profiling to directly probe the function of all single copy sequences of S. pombe. As a proof of principle, we used integration profiling to determine which genes are required for cells to grow in defined medium. We generated a large library of Hermes integration events and sequenced 360,000 independent insertions, or one integration for every 29 nt of the genome. Approximately 33% of insertions occurred in ORFs. ORFs fell into two classes, with one group having significantly higher densities of integration than the other. Using essentiality designations from the Bioneer Deletion Consortium, we found that ORFs with higher densities of integration corresponded to nonessential genes while the ORFs with lower numbers of insertions corresponded to essential genes. We also studied the relationship of Hermes integration and chromatin structures and found that Hermes has a distinct preference for integrating in-between nucleosomes. This study highlights the powerful use of integration profiling to create a genome-wide footprint of essential sequences in S. pombe.

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