http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2019136169-A1

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filingDate 2019-01-03-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_6703040485afca166ab0a669dcf52090
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publicationDate 2019-07-11-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber WO-2019136169-A1
titleOfInvention Versatile amplicon single-cell droplet sequencing-based shotgun screening platform to accelerate functional genomics
abstract Disclosed is a method of functional genomics determination including transducing a cell population with a set of nucleic acid molecules including a pooled library of genomic perturbagens to integrate multiple perturbagen cassettes into the genome. A phenotype of individual cells is determined and single cells of the population with targeted phenotypes are individually sorted into a set of compartments. Each compartment includes a forward primer with a nucleic acid sequence (NAS) that specifically binds a common nucleic acid sequence on the nucleic acid molecules and a compartment (cell)-specific nucleic acid barcode. Also included is a reverse primer with a NAS that specifically binds a common NAS on the nucleic acid molecules comprising a pooled library of genomic perturbagens. The genome-integrated perturbagen cassettes are create amplicons which are pooled and sequences determined. This method can be applied to other genome-level single-cell applications - immune receptor profiling, targeted DNA/RNA sequencing, and metagenomics.
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