An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting

Cao, Jian; Wu, Lizhen; Zhang, Shang-Min; Lü, Min; Cheung, William K.C.; Cai, Wesley L.; Gale, Molly; Xu, Qi; Yan, Qin

doi:10.1093/nar/gkw660

Cited by 164 publications

(166 citation statements)

References 50 publications

(74 reference statements)

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“…as well as the doxycycline-inducible Cas9 viral vectors. 7 Indeed, considerably decreased off-target effects without influence on on-target efficiency have been observed using these systems, a finding that could support the working strategy that Cas9 expression duration can be manipulated to achieve higher specificity. However, there is potential toxicity associated with small molecular inducers.…”

Section: A Self-restricted Crispr System To Reduce Offtarget Effectsmentioning

confidence: 74%

A Self-restricted CRISPR System to Reduce Off-target Effects

et al. 2016

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Section: A Self-restricted Crispr System To Reduce Offtarget Effectsmentioning

confidence: 74%

A Self-restricted CRISPR System to Reduce Off-target Effects

et al. 2016

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“…48h later, cells were selected by treating with puromycin (2ug/ml) for 2 weeks and continually cultured in puromycin afterwards. The knockout efficiency was validated by western blot [18]. …”

Section: Methodsmentioning

confidence: 99%

B Subunit of Human Chorionic Gonadotropin Promotes Tumor Invasion and Predicts Poor Prognosis of Early-Stage Colorectal Cancer

Yin

Song

et al. 2018

Cell Physiol Biochem

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Background/Aims: It is well established that many non-trophoblastic tumors secrete HCG (human chorionic gonadotropin) and that such secretion is correlated with the poor prognosis of tumor patients. This study aims to analyze the correlation between β-HCG expression and outcome of colorectal cancer (CRC) and understand its role in CRC pathology Methods: We detected the mRNA and protein expression of β-HCG in human CRC tissues with RT-qPCR and immunohistochemistry, and we compared the clinical-pathological characteristics, prognosis and progression between the β-HCG positive and negative groups. We also generated CRC cell lines with β-HCG over-expression as well as β-HCG stable knockout, and evaluated cell function and mechanism in vitro and in vivo. Results: Fifty out of 136 CRC patients (37%) expressed β-HCG at the invasive front. Clinical-pathological data showed that β-HCG was positively correlated with Dukes staging (P=0.031) and lymph node metastasis (P=0.012). Survival analysis suggested that the patients with high expression of β-HCG had poorer prognosis than those with low β-HCG expression (P=0.0289). β-HCG expression level was also positively correlated with tumor invasion in early-stage CRC patient tissues (P=0.0227). Additionally β-HCG promoted the migration and invasion of CRC in vitro and in vivo but had no effect on the proliferation of tumor cells. Conclusion: Our study demonstrated that β-HCG was ectopically expressed in the CRC patients and its high expression correlated with poor prognosis of early-stage CRC. Additionally it worked as an oncogene that promotes the migration and invasion of CRC by epithelial-mesenchymal transition (EMT).

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“…Physicians and patients alike are certainly right to seek to the medical risks of vaccines, such as allergies to certain ingredients like eggs or latex,65 and the rarer risks of anaphylaxis and encephalopathy;51 likewise, they must consider the predominant medical risk of CRISPR––off-target effects66 67––which scientists are working to diminish 68. However, to allow fear of these risks to obfuscate these innovations' very tangible benefits would be an irresponsible and potentially more dangerous option.…”

Section: Resultsmentioning

confidence: 99%

Everything in moderation, even hype: learning from vaccine controversies to strike a balance with CRISPR

Benston

2017

J Med Ethics

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The ease and applicability of CRISPR/Cas9—a new and precise gene-editing and reproductive technology—have garnered hype and heightened concern about its potential “unprecedented and horrific consequences” and have led many scientific leaders to call for a moratorium on its research and use. CRISPR appears distinctly more controversial than previous technological innovations (genetic or otherwise), with a greater reach and speed of human treatment and enhancement; however, we have seen similarly inflated hopes and fears in response to other medical innovations for well over a century. One intervention that has both historically and recently incited alarm—vaccines—serves as a pertinent example of what could go wrong if a technology’s reach is shortened due to inflated fears. By comparing the vaccine controversy and the CRISPR debate, we can help separate the hype from the realistic potential of these technologies. How our society grapples with such innovations will determine the extent to which their impact on our individual and collective health will be beneficial. We must recognize the need for a tempered approach to CRISPR conversation leading to regulation and ethical application. Although CRISPR’s reach will continue expanding with ongoing research, thus requiring continuous evaluation, the lessons we’ve learned from the vaccine controversy demonstrate that our approach must not be to shut down regulation and application now, but to thoughtfully conjoin productive debate and action so that therapeutic gene-editing can alleviate suffering as soon as possible without precipitating social outcomes we would belatedly deplore.

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An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting

Cited by 164 publications

References 50 publications

A Self-restricted CRISPR System to Reduce Off-target Effects

A Self-restricted CRISPR System to Reduce Off-target Effects

B Subunit of Human Chorionic Gonadotropin Promotes Tumor Invasion and Predicts Poor Prognosis of Early-Stage Colorectal Cancer

Everything in moderation, even hype: learning from vaccine controversies to strike a balance with CRISPR

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