Unexpected mutations after CRISPR–Cas9 editing in vivo

Schaefer, Kellie A.; Wu, W. D.; Colgan, Diana F.; Tsang, Stephen H.; Bassuk, Alexander G.; Mahajan, Vinit B.

doi:10.1038/nmeth.4293

Cited by 306 publications

(251 citation statements)

References 6 publications

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“…19 Off-target analysis was a secondary outcome reported in our Correspondence. 1 In our study, only two of the eleven founders showed successful HDR. 19 Tissue from these two and a colony control underwent WGS.…”

Section: Identity Of Some Mutations In Casesmentioning

confidence: 52%

Corrigendum and follow-up: Whole genome sequencing of multiple CRISPR-edited mouse lines suggests no excess mutations

Schaefer

Darbro

Colgan

et al. 2017

Preprint

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Our previous publication suggested CRISPR-Cas9 editing at the zygotic stage might unexpectedly introduce a multitude of subtle but unintended mutations, an interpretation that not surprisingly raised numerous questions. The key issue is that since parental lines were not available, might the reported variants have been inherited? To expand upon the limited available whole genome data on whether CRISPR-edited mice show more genetic variation, whole-genome sequencing was performed on two other mouse lines that had undergone a CRISPR-editing procedure. Again, parents were not available for either the Capn5 nor Fblim1 CRISPR-edited mouse lines, so strain controls were examined. Additionally, we also include verification of variants detected in the initial mouse line. Taken together, these whole-genome-sequencing-level results support the idea that in specific cases, CRISPR-Cas9 editing can precisely edit the genome at the organismal level and may not introduce numerous, unintended, off-target mutations.

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Section: Identity Of Some Mutations In Casesmentioning

confidence: 52%

Corrigendum and follow-up: Whole genome sequencing of multiple CRISPR-edited mouse lines suggests no excess mutations

Schaefer

Darbro

Colgan

et al. 2017

Preprint

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show abstract

“…c Genetic engineering in rice using dual gRNAs targeting ESPS introns for double amino acid substitution [T102I + P106S (TIPS)]. d Systematic testing of enzyme perturbation sensitivities (STEPS) approach to iteratively find bottleneck ◂ technology, with several studies pointing out unwanted cuts due to the gRNA binding elsewhere than the intended target region [27,85]. This phenomenon is known to be accentuated in regions with sequences similar to the original sequence, e.g.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

Ferreira

David

Nielsen

2018

Journal of Industrial Microbiology and Biotechnology

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Clustered regularly interspaced short palindromic repeats (CRISPR) is poised to become one of the key scientific discoveries of the twenty-first century. Originating from prokaryotic and archaeal immune systems to counter phage invasions, CRISPRbased applications have been tailored for manipulating a broad range of living organisms. From the different elucidated types of CRISPR mechanisms, the type II system adapted from Streptococcus pyogenes has been the most exploited as a tool for genome engineering and gene regulation. In this review, we describe the different applications of CRISPR/Cas9 technology in the industrial biotechnology field. Next, we detail the current status of the patent landscape, highlighting its exploitation through different companies, and conclude with future perspectives of this technology.

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“…Due to the cleavage activation of Cas9 which can mutate the homologous host genome, the risk of molecular cocktail seems to be inevitable and the side effect is uncertain. Researchers found that CRISPR/Cas9 which was used to edit a gene of blindness in the mice could lead to hundreds of unexpected mutations [46]. Then, in the process of screening and culturing the cells, there are also problems.…”

Section: The Challenges and Opportunitiesmentioning

confidence: 99%

CRISPRs encounter Cocktail: a dawn for curing HIV/AIDS

Li¹,

Zhang²,

Feng³

et al. 2017

Virol Res Rev

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a These authors contributed equally to this work. AbstractThe acquired immunodeficiency syndrome (AIDS) patients can scarcely be cured completely because of the Human Immunodeficiency Virus (HIV) provirus existence post-infection in body, though cocktail of the highly active antiretroviral therapy (HAART) has achieved great effects on controlling and decreasing HIV clinically. Recently, the genome editing strategies are developing by leaps and bounds. Among three generation of technologies, the clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein 9 (Cas9) is a newborn to former zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) but grows fast into the most popular at present, because of its advantages in less time-and cost-consuming. Eradicating HIV by disrupting the viral co-receptor C-C chemokine receptor type five (CCR5) or C-X-C chemokine receptor type four (CXCR4) gene, excising the provirus segments integrated into human genome, or activating/inactivating the replication of the virus genome by using these technologies are several current strategies to reach the goal of AIDS prevention and treatment. It seems hard, however, to arrive the keen anticipation exactly by using them respectively. After a comprehensive literature review, it concluded that the combination of those interventions, as together co-receptor with provirus genome interference as a cocktail edition, may lead us to an eventual cure for the horrible disease.

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Unexpected mutations after CRISPR–Cas9 editing in vivo

Cited by 306 publications

References 6 publications

Corrigendum and follow-up: Whole genome sequencing of multiple CRISPR-edited mouse lines suggests no excess mutations

Corrigendum and follow-up: Whole genome sequencing of multiple CRISPR-edited mouse lines suggests no excess mutations

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

CRISPRs encounter Cocktail: a dawn for curing HIV/AIDS

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