AI Article Synopsis
- The CRISPR/Cas9 system is a groundbreaking technology for modifying genomes using the bacterial enzyme Cas9 and a custom guide RNA (gRNA).
- This method allows for the precise cutting of DNA at specific locations, enabling targeted deletions of DNA segments in human chromosomes.
- The technique can effectively delete genomic regions from several hundred base pairs up to 1 million base pairs, with a deletion efficiency of 1-10%, making it potentially valuable for creating modified cell and animal models.
Article Abstract
The CRISPR/Cas9 system has emerged as an intriguing new technology for genome engineering. It utilizes the bacterial endonuclease Cas9 which, when delivered to eukaryotic cells in conjunction with a user-specified small guide RNA (gRNA), cleaves the chromosomal DNA at the target site. Here we show that concurrent delivery of gRNAs designed to target two different sites in a human chromosome introduce DNA double-strand breaks in the chromosome and give rise to targeted deletions of the intervening genomic segment. Predetermined genomic DNA segments ranging from several-hundred base pairs to 1 Mbp can be precisely deleted at frequencies of 1-10%, with no apparent correlation between the size of the deleted fragment and the deletion frequency. The high efficiency of this technique holds promise for large genomic deletions that could be useful in generation of cell and animal models with engineered chromosomes.
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| http://dx.doi.org/10.1002/bit.25490 | DOI Listing |
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