FIELD: biotechnology.
SUBSTANCE: proposed double-stranded ring polynucleotide comprises an exogenous nucleic acid sequence and two sequences flanking said exogenous sequence. In this case, flanking sequences contain two or more target regions for two or more naturally occurring nucleases. These target sites flank a 4–20 nucleotide spacer sequence that does not occur in the genome of said cell. Proposed double-stranded polynucleotide does not contain homology arms. Proposed polynucleotide and method can be used in biotechnology for directional insertion of donor molecules independent of homology into the genome of the cell.
EFFECT: annular double-stranded polynucleotide was proposed to integrate exogenous nucleic acid into the genome of the cell and method for integrating an exogenous nucleic acid sequence into an endogenous cell locus.
16 cl, 61 dwg, 33 tbl, 10 ex
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