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1. (WO2017156336) TRANSPOSASE-MEDIATED IMAGING OF THE ACCESSIBLE GENOME

Pub. No.:    WO/2017/156336    International Application No.:    PCT/US2017/021677
Publication Date: Fri Sep 15 01:59:59 CEST 2017 International Filing Date: Fri Mar 10 00:59:59 CET 2017
IPC: C12N 9/22
C12N 15/11
C12N 15/87
C12Q 1/68
Applicants: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Inventors: CHANG, Howard Y.
GREENLEAF, William J.
CHEN, Xingqi
BUENROSTRO, Jason
Title: TRANSPOSASE-MEDIATED IMAGING OF THE ACCESSIBLE GENOME
Abstract:
Methods for labeling and imaging the accessible genome using a transposase are disclosed. In some embodiments, a bifunctional transposase complex or transposome is used to insert adaptors comprising chemical tags selectively at accessible sites in the genome where active regulatory DNA is located. Various chemical tags can be used for labeling DNA at insertion sites, including, for example, fluorescent dyes for fluorescence imaging, metal particles for electron microscopy or magnetic manipulation of DNA, isotopic labels, or biotin or other ligands, haptens, substrates, or inhibitors that are recognized by streptavidin, antibodies, enzymes, or receptors. Labeling DNA in this manner can be used to provide spatial information regarding the positioning of regulatory DNA in the genome and makes possible the imaging and sorting of cells based on the status of their regulatory DNA.