公开(公告)号：US20210292752A1

公开(公告)日：2021-09-23

申请号：US17266566

申请日：2019-08-13

Applicant: Spiber Inc. ,  The University of Tokyo

Inventor： Kana Ishida ,  Soh Ishiguro ,  Nozomu Yachie ,  Chikako Sato ,  Junichi Sugahara

IPC: C12N15/10

Abstract: Disclosed is a method for isolating or identifying target clone cells from a cell population, the method including steps of: preparing a cell population into which a barcode sequence and at least one reporter protein abnormal expression cassette linked to the barcode sequence are introduced; introducing a barcode sequence recognition module targeting an arbitrary barcode sequence and a nucleic acid mutation repair enzyme into cells; repairing a nucleic acid mutation causing abnormal expression occurring in the at least one reporter protein abnormal expression cassette by expression of a complex of the barcode sequence recognition module and the nucleic acid mutation repair enzyme in a cell containing the target barcode sequence, to induce normal expression of the reporter protein; and isolating or identifying target clone cells in which the reporter protein is expressed.

公开(公告)号：US20200248231A1

公开(公告)日：2020-08-06

申请号：US16641946

申请日：2018-08-28

Applicant: Spiber Inc.

Inventor： Yusuke Ito ,  Chikako Sato ,  Nozomu Yachie

IPC: C12Q1/686

Abstract: The present invention relates to a method for producing a double-stranded DNA fragment having a desired nucleotide sequence through dual asymmetric PCR (DA-PCR). The method comprises: (1) providing a plurality of oligonucleotides (sense oligonucleotides) each corresponding to a part of a sense strand of the double-stranded DNA fragment and a plurality of oligonucleotides (antisense oligonucleotides) each corresponding to a part of an antisense strand of the double-stranded DNA fragment and mixing together the oligonucleotides with equal concentrations, DNA polymerase, and dNTP to prepare a reaction mixture solution; (2) performing PCR by using the reaction mixture solution from step (1); (3) adding a primer set capable of amplifying the double-stranded DNA fragment of full length to the reaction mixture solution from step (2); and (4) performing PCR by using the reaction mixture solution from step (3).