SCREENING OF CAS NUCLEASES FOR ALTERED NUCLEASE ACTIVITY
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United States of America Patent
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Jan 23, 2025
app pub date -
Nov 15, 2022
filing date -
Nov 15, 2022
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Described herein is a method of screening for a nuclease with an altered nuclease activity comprising; a) forming a first compartmentalizing reaction comprising a carrier and a variant nuclease template nucleic acid comprising a coding region for a variant nuclease, and amplifying said coding region for said variant nuclease, to obtain variant nuclease encoding nucleic acid; b) forming a second compartmentalizing reaction comprising said variant nuclease encoding nucleic acid, and performing an in vitro transcription and translation reaction to obtain variant nuclease polypeptides; and c) forming a third compartmentalizing reaction comprising said variant nuclease polypeptides, and assaying said variant nuclease polypeptides for the altered nuclease activity of said nuclease.
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