BioTechniques
 2014
DOI: 10.2144/000114196
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Precise Gene Deletion and Replacement Using the CRISPR/Cas9 System in Human Cells

Qiupeng Zheng
1
,
Xiaohong Cai
2
,
Meng How Tan
3
et al.

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 151 publications

(97 citation statements)
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“…1e and Supplementary Fig. 1e), consistent with previous findings 18 . Taken together, lentivirally-delivered pgRNAs are capable of creating large genomic deletions with high efficiency in mammalian cells.…”
Section: Resultssupporting
confidence: 92%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
show abstract
“…1e and Supplementary Fig. 1e), consistent with previous findings 18 . Taken together, lentivirally-delivered pgRNAs are capable of creating large genomic deletions with high efficiency in mammalian cells.…”
Section: Resultssupporting
confidence: 92%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
show abstract
“…10,31 In this study, two different guides were used simultaneously with the aim of deleting the intermediate sequence and restricting the range of genome modifications, as has been previously indicated. [12][13][14] However, it did not appear as a solution in zygotes, which is contrary to what has been described in adult cells. 12,32,33 The preferential pathway for DNA repair differs between embryonic and adult cells and, in addition, DNA repair is accelerated in embryonic cells compared with adult cells.…”
Section: Discussioncontrasting
confidence: 86%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
show abstract
“…As mentioned above, these additional deletions can produce predictably four types of results. Indeed, our results confirmed that gRNAs and Cas9 cut DNA most of the times at exactly three nucleotides from the PAM in the 5′ direction 45 and produced the predicted results.…”
Section: Discussionsupporting
confidence: 83%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
show abstract
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