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Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one-cell mouse embryos.
FEBS Open Bio 5, 26-35 (2015)
The use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one-cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant double-strand breaks. Herein, we applied pairs of TALEN or sgRNAs and Cas9 to create deletions in the Rab38 gene. We found that the deletion of 3.2 or 9.3. kb, but not of 30. kb, occurs at a frequency of 6-37%. This is sufficient for the direct production of mutants by embryo microinjection. Therefore, deletions up to ~10. kb can be readily achieved for modeling human disease alleles. This work represents an important step towards the establishment of new protocols that support the ligation of remote DSB ends to achieve even larger rearrangements.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Cas9 ; Crispr ; Disease Model ; Mouse Mutant ; One-cell Embryo ; Talens
ISSN (print) / ISBN
2211-5463
Journal
FEBS Open Bio
Quellenangaben
Volume: 5,
Pages: 26-35
Publisher
Elsevier
Publishing Place
Cambridge
Non-patent literature
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Reviewing status
Peer reviewed