Engineered secreted reporters of cellular processes
OwnerAryeh Warmflash, Kinshuk Mitrainventors
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Abstract
A method of determining an amount of transcript in a living organism, the method requiring editing a genome of a living organism to include a DNA copy of a unique microRNA barcode downstream of a promoter of a transcript to be measured, such that expression of that transcript includes expression of said microRNA barcode. Then, one collects secretions from said living organism and measures a level of said transcript by measuring the amount of the unique microRNA barcode present in those secretions. Cells, tissue and animals containing such barcodes are also included.
Claims19
A method of determining an amount of transcript in a living organism, said method comprising:
- a) editing a genome of a living organism to contain a unique microRNA barcode;
- b) collecting secretions from said living organism that includes a DNA copy of said unique microRNA barcode downstream of a promoter of a transcript to be measured, such that expression of said transcript includes expression of said unique microRNA barcode;
- and
- c) measuring a level of said transcript by measuring an amount of said unique microRNA barcode in said secretions.
A method of determining an amount of transcript in a living organism, said method comprising:
- a) editing a genome of a living organism to insert a DNA copy of a synthetic intron comprising a unique microRNA barcode with less than 15% homology to said genome downstream of a promoter of a transcript to be measured, such that expression of said transcript includes expression of said microRNA barcode, wherein said microRNA barcode is nonfunctional and is not intended to modulate expression of any gene in said living organism;
- b) extracting secretions from said living organism;
- c) purifying RNA from said secretions;
- d) converting said microRNA barcodes in said RNA to DNA barcodes and amplifying said DNA barcodes;
- and,
- e) measuring a level of said transcript in said living organism by measuring an amount of said amplified DNA barcode.
Dependencies are inferred from the claim text.