BASE EDITOR, PREPARATION METHOD AND USE THEREOF

BASE EDITOR, PREPARATION METHOD AND USE THEREOF

The present invention provides a base editor, a preparation method and use thereof, and belongs to the technical field of gene editing. The base editor includes a pCMV-dCpfl-RR-eBE recombinant plasmid and a pLbCpfl-sgRNA recombinant plasmid. The use includes the following steps: determining a target...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Hauptverfasser: Li, Hegang, Xiaojing, Hao, Zhao, Jinshan, Qin, Huaiyuan, Zhang, Ning, Xin, Jingjing
Format: Patent
Sprache:eng
Schlagworte:
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
Online-Zugang:Volltext bestellen
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung:The present invention provides a base editor, a preparation method and use thereof, and belongs to the technical field of gene editing. The base editor includes a pCMV-dCpfl-RR-eBE recombinant plasmid and a pLbCpfl-sgRNA recombinant plasmid. The use includes the following steps: determining a target sequence and designing a single-stranded oligonucleotide pair; annealing to obtain a double-stranded DNA fragment; ligating to the pLbCpfl-sgRNA recombinant plasmid to obtain a targeting sgRNA expression vector; and culturing after cotransfection of cells with the targeting sgRNA expression vector and the pCMV-dCpfl-RR-eBE recombinant plasmid. The base editor of the present invention specifically mutates cytosine (C) to thymine (T) at a target site, but has no effect on bases at a non-target site. Gene editing efficiency ranges between 20% and 30%.