2014
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Abstract: The prokaryotic type II CRISPR/Cas9 system has been adapted to perform targeted genome editing in cells and model organisms. Here, we describe targeted gene deletion and replacement in human cells via the CRISPR/Cas9 system using two guide RNAs. The system effectively generated targeted deletions of varied length, regardless of the transcriptional status of the target gene. It is notable that targeted gene deletions generated via CRISPR/Cas9 and two guide RNAs resulted in the formation of correct junctions at … Show more

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Cited by 119 publications
(73 citation statements)
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References 23 publications
(73 reference statements)
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“…Use of two gRNAs coupled with Cas9 can efficiently create DNA deletions of up to 10 kb in the presence of a linear homologous repair donor [108]. Interrogation of gene function on a genome-wide scale can be facilitated by large scale oligonucleotide synthesis of guide sequences [109].…”
Section: Functional Genomics By Crispr-cas9mentioning
confidence: 99%
“…Use of two gRNAs coupled with Cas9 can efficiently create DNA deletions of up to 10 kb in the presence of a linear homologous repair donor [108]. Interrogation of gene function on a genome-wide scale can be facilitated by large scale oligonucleotide synthesis of guide sequences [109].…”
Section: Functional Genomics By Crispr-cas9mentioning
confidence: 99%
“…2A). Targeting a cellular gene with two gRNA/Cas9 complexes will result, in many cases, in a specific gene fragment deletion (10). To mutate the cGAS and STING genes, two gRNA sequences per gene were designed to delete a 32-bp (cGAS) and 149-bp (STING) fragment just downstream of the translation initiation site (Fig.…”
mentioning
confidence: 99%
“…Four out of six (M2_9, M2_10, M3_10 and M3_11) showed the expected 37 nt ‘clean’ deletion without any additional mutations. Precise deletions, such as this, using 2 sgRNAs have previously been generated with high efficiency [37, 42], and allow a large degree of control over the mutation. Two of the sub-clones (M3_9 and M2_12) showed one allele with the expected 37 nt deletion and the other with an additional deletion at the 2nd sgRNA cut site.…”
Section: Resultsmentioning
confidence: 99%
“…Precise deletions larger than a gene using CRISPR-Cas and two sgRNAs have been previously demonstrated [42]. …”
Section: Resultsmentioning
confidence: 99%