RRC ID 33279
Author Kaneko T, Sakuma T, Yamamoto T, Mashimo T.
Title Simple knockout by electroporation of engineered endonucleases into intact rat embryos.
Journal Sci Rep
Abstract Engineered endonucleases, such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, provide a powerful approach for genome editing in animals. However, the microinjection of endonucleases into embryos requires a high skill level, is time consuming, and may cause damage to embryos. Here, we demonstrate that the electroporation of endonuclease mRNAs into intact embryos can induce editing at targeted loci and efficiently produce knockout rats. It is noteworthy that the electroporation of ZFNs resulted in an embryonic survival rate (91%) and a genome-editing rate (73%) that were more than 2-fold higher than the corresponding rates from conventional microinjection. Electroporation technology provides a simple and effective method to produce knockout animals.
Volume 4
Pages 6382
Published 2014-10-1
DOI 10.1038/srep06382
PII srep06382
PMID 25269785
PMC PMC4180828
MeSH Animals CRISPR-Cas Systems* Cells, Cultured Clustered Regularly Interspaced Short Palindromic Repeats / genetics* Electroporation / methods* Embryo Transfer Embryo, Mammalian / cytology Embryo, Mammalian / metabolism* Endodeoxyribonucleases / genetics Endodeoxyribonucleases / metabolism* Female Gene Targeting / methods* Genetic Engineering / methods* Male Rats Rats, Inbred F344 Zinc Fingers
IF 3.998
Times Cited 82
Rats F344/Stm(strainID=266)
Human and Animal Cells Rat-1(RCB1830)