AI Article Synopsis
- - CRISPR technology allows for easy creation of knockout and knock-in animal models, but these models may have unintended off-target effects or random DNA insertions.
- - Current methods to detect these issues, such as off-target lesions and random insertions, are limited and often not straightforward.
- - The newly developed method, CRISPR-KRISPR, can efficiently identify off-target effects and random insertions using minimal genomic DNA, making it a practical addition for screening founder animals in breeding programs.
Article Abstract
CRISPR tools can generate knockout and knock-in animal models easily, but the models can contain off-target genomic lesions or random insertions of donor DNAs. Simpler methods to identify off-target lesions and random insertions, using tail or earpiece DNA, are unavailable. We develop CRISPR-KRISPR (CRISPR-Knock-ins and Random Inserts Searching PRotocol), a method to identify both off-target lesions and random insertions. CRISPR-KRISPR uses as little as 3.4 μg of genomic DNA; thus, it can be easily incorporated as an additional step to genotype founder animals for further breeding.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9594901 | PMC |
| http://dx.doi.org/10.1186/s13059-022-02779-8 | DOI Listing |
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