Method for identifying functional elements
AssigneesPeking University, Edigene Inc.
InventorsWensheng WEI, Yinan WANG, Yuexin ZHOU, Xinyi ZHANG, Di YUE, Ying LIU
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Abstract
Provided are a method for identifying functional elements of a genomic sequence and a library used for identifying functional elements of a genomic sequence.
Claims40
A library used for identifying functional elements of a genomic sequence comprising a plurality of CRISPR-Cas system guide RNAs comprising guide sequences that are capable of targeting a plurality of genomic sequences within at least one continuous genomic region, wherein the guide RNAs target at least 100 genomic sequences comprising non-overlapping cleavage sites upstream of a PAM sequence for every 1000 base pairs within the continuous genomic region.
A method for identifying functional elements for a protein of interest comprising conducting saturation mutagenesis to the protein of interest by disrupting the genomic gene coding for the protein by using CRISPR-Cas system introduced into a population of cells, determining disrupted genomic sites associated with change of phenotype by sequencing DNA and cDNA of the targeted gene, retrieving in-frame mutations that give rise to the change of phenotype, and building a bioinformatics pipeline to identify functional elements of the protein of interest at single amino acid resolution.
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Dependencies are inferred from the claim text.