GENE EDITING USING MODIFIED CLOSED-END DNA (ceDNA) Russian patent published in 2024 - IPC C12N5/10 C12N15/09 C12N15/63 C12N15/64 

FIELD: biotechnology; genetic engineering.

SUBSTANCE: invention is a method of inserting a donor sequence into a predetermined insertion site on a chromosome in a host cell, comprising: introducing a guide RNA (gRNA) and an mRNA encoding a Cas enzyme into the host cell; and a composition comprising a lipid nanoparticle (LNP) containing a capsid-less closed-end DNA vector (ceDNA); wherein the ceDNA vector contains a nucleotide sequence between flanking inverted terminal repeats (ITR), wherein at least one nucleotide sequence is a gene-editing nucleic acid sequence containing a 5'-homologous arm, a repair template donor sequence and a 3'-homologous arm, and wherein the donor sequence is inserted into the chromosome at or near said insertion site by homologous recombination. The invention also relates to a method for editing chromosomal genes in a host cell, including introducing into the host cell a guide RNA (gRNA) and an mRNA encoding the Cas enzyme; and a composition comprising a lipid nanoparticle (LNP) containing a capsid-less closed-end DNA vector (ceDNA); wherein the ceDNA vector contains a nucleotide sequence between flanking inverted terminal repeats (ITRs), wherein at least one nucleotide sequence is a gene-editing nucleic acid sequence, wherein the gene-editing nucleic acid sequence contains a donor sequence, wherein said chromosome is restored using non-homologous end joining (NHEJ).

EFFECT: invention makes it possible to effectively insert a donor sequence into a certain part of the host cell chromosome.

62 cl, 19 dwg, 10 tbl, 18 ex