method for producing rna multiplex-mediated genetic manipulation in a receptor cell, nucleic acid construct, expression cassette, vector, receptor cell, genetically modified cell, tissue, plant or animal, plant or animal progeny, and method for modifying a target site in the genome of a cell

摘要

The invention includes materials and methods for generating numerous small RNAs from a polynucleotide (synthetic gene) construct to facilitate the RNA-guided multiplex reaction in genome editing, modification, expression inhibition, and other RNA-based technologies. The synthetic gene / polynucleotide construct encodes the RNA-separated polycistronic RNA components and preferably also includes regulatory components such as a promoter or terminator to form an expression cassette. once transcribed into a cell, the transcript is processed by the cell to multiple rna molecules by the endogenous rnat processing system. the system can be applied to any rna-based gene manipulation method, including rna-mediated genome editing, artificial microrna-mediated gene silencing, small-rna-mediated rna genetic manipulation, double-stranded rna gene silencing, antisense mechanisms and the like.