DESIGNING SYSTEMS, METHODS AND OPTIMIZED GUIDE COMPOSITIONS FOR MANIPULATING SEQUENCES Russian patent published in 2019 - IPC C12N15/63 

FIELD: biotechnologies.

SUBSTANCE: invention relates to the biotechnology. Described is a CRISPR-Cas system for editing a genome in a eukaryotic cell, comprising: Cas9 protein containing at least one nuclear localization sequence, and a chimeric RNA (chiRNA) of the CRISPR-Cas system, comprising: (a) a guide sequence capable of hybridising with a target sequence in a eukaryotic cell, (b) a paired tracr sequence capable of hybridising with the tracr-sequence, and (c) tracr-sequence, where (a), (b) and (c) are located in 5'-3' orientation, where one or more of the tracr- and paired tracr-sequence guides are modified to increase stability and where optionally a protein Cas9 forms a complex with chimeric RNA (chiRNA) of the CRISPR-Cas system. Also presented is the CRISPR-Cas vector system for modifying the target sequence in a eukaryotic cell containing one or more vectors containing: I. a first regulatory element operably linked to a nucleotide sequence encoding a chimeric RNA (chiRNA) of the CRISPR-Cas system, comprising: (a) a guide sequence capable of hybridising with a target sequence in a eukaryotic cell, (b) a paired tracr sequence capable of hybridising with a tracr sequence, and (c) a tracr sequence, where (a), (b) and (c) are located in 5'-3' orientation, and II. a second regulatory element operably linked to a nucleotide sequence encoding a Cas9 protein comprising at least one nuclear localization sequence, where components I and II are in the same or different vectors of the system, where one or more of the tracr- and paired tracr-sequences are modified to increase stability.

EFFECT: invention extends the range of products which control expression.

49 cl, 23 dwg, 4 tbl, 8 ex