Drug-tunable multidimensional synthetic gene control using inducible degron-tagged dCas9 effectors

Kleinjan, Dirk A.; Wardrope, Caroline; Sou, Si Nga; Rosser, Susan J.

doi:10.1038/s41467-017-01222-y

Cited by 60 publications

(48 citation statements)

References 21 publications

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“…Similarly, this e ffect was rapidly reversed when auxin was removed from the media, leading to the reappearance of detectable levels of the fusion protein in 30 min and maximum expression in the next 3−7 h. The AID approach was also extended to specific orthogonal Cas9 and Cpf1 proteins, a ffording orthogonal, auxin-degradable synthetic transcription factors. 87 Jacobs et al inserted stabilizable polypeptide linkages (StaPLs) based on hepatitis C virus NS3 protease domain into internal loops of dCas9 to enable conditional control of Cas9 activity. StaPLs undergo autoproteolysis to cleave the proteins to which they are attached, though the presence of a protease inhibitor prevents their cleavage to preserve protein function.…”

Section: Control By Small-molecule Activatorsmentioning

confidence: 99%

Precision Control of CRISPR-Cas9 Using Small Molecules and Light

et al. 2019

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The CRISPR (clustered regularly interspaced short palindromic repeat)-Cas system is an adaptive immune system of bacteria that has furnished several RNA-guided DNA endonucleases (e.g., Cas9) that are revolutionizing the field of genome engineering. Cas9 is being used to effect genomic alterations as well as in gene drives, where a particular trait may be propagated through a targeted species population over several generations. The ease of targeting catalytically impaired Cas9 to any genomic loci has led to development of technologies for base editing, chromatin imaging and modeling, epigenetic editing, and gene regulation. Unsurprisingly, Cas9 is being developed for numerous applications in biotechnology and biomedical research and as a gene therapy agent for multiple pathologies. There is a need for precise control of Cas9 activity over several dimensions, including those of dose, time, and space in these applications. Such precision controls, which are required of therapeutic agents, are particularly important for Cas9 as off-target effects, chromosomal translocations, immunogenic response, genotoxicity, and embryonic mosaicism are observed at elevated levels and with prolonged activity of Cas9. Here, we provide a perspective on advances in the precision control of Cas9 over aforementioned dimensions using external stimuli (e.g., small molecules or light) for controlled activation, inhibition, or degradation of Cas9.

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Section: Control By Small-molecule Activatorsmentioning

confidence: 99%

Precision Control of CRISPR-Cas9 Using Small Molecules and Light

et al. 2019

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“…It remains unknown whether other cytidine deaminases will cause similar off-target deamination (Kim et al 2017b;Koblan et al 2018). In addition, it might be feasible to reduce CBE off-target effects by delivering ribonucleoprotein instead of DNA or RNA, controlling CBE expression and duration by inducible system (Rees et al 2017;Dow et al 2015;Kleinjan et al 2017). On the other hand, CBE off-target effects should be studied in a Breal-world^preclinical application, such as human somatic cells.…”

mentioning

confidence: 99%

Off-target challenge for base editor-mediated genome editing

Liang

Huang

2019

Cell Biol Toxicol

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“…Lentiviral integration produces stable, constitutive expression of CRISPR components, which could potentially be undesirable in the long term, for example by increasing off‐target effects of CRISPR nucleases. However, lentiviral delivery systems can be adapted to limit the window of Cas protein activity by using inducible promoters, or conditionally degradable or stabilizable Cas proteins …”

Section: Challenges In Applying Crispr/cas Gene Editing For Respiratomentioning

confidence: 99%

Applications of CRISPR systems in respiratory health: Entering a new ‘red pen’ era in genome editing

Moses

Kaur

2019

Respirology

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Respiratory diseases, such as influenza infection, acute tracheal bronchitis, pneumonia, tuberculosis, chronic obstructive pulmonary disease, asthma, lung cancer and nasopharyngeal carcinoma, continue to significantly impact human health. Diseases of the lung and respiratory tract are influenced by environmental conditions and socio-economic factors; however, many of these serious respiratory disorders are also rooted in genetic or epigenetic causes. Clustered regularly interspaced palindromic repeats (CRISPR) and CRISPRassociated (Cas) proteins, isolated from the immune system of prokaryotes, provide a tool to manipulate gene sequences and gene expression with significant implications for respiratory research. CRISPR/Cas systems allow preclinical modelling of causal factors involved in many respiratory diseases, providing new insights into their underlying mechanisms. CRISPR can also be used to screen for genes involved in respiratory processes, development and pathology, identifying novel disease drivers or drug targets. Finally, CRISPR/-Cas systems can potentially correct genetic mutations and edit epigenetic marks that contribute to respiratory disorders, providing a form of personalized medicine that could be used in conjunction with other technologies such as stem cell reprogramming and transplantation. CRISPR gene editing is a young field of research, and concerns regarding its specificity, as well as the need for efficient and safe delivery methods, need to be addressed further. However, CRISPR/Cas systems represent a significant step forward for research and therapy in respiratory health, and it is likely we will see the breakthroughs generated from this technology continue.

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Drug-tunable multidimensional synthetic gene control using inducible degron-tagged dCas9 effectors

Cited by 60 publications

References 21 publications

Precision Control of CRISPR-Cas9 Using Small Molecules and Light

Precision Control of CRISPR-Cas9 Using Small Molecules and Light

Off-target challenge for base editor-mediated genome editing

Applications of CRISPR systems in respiratory health: Entering a new ‘red pen’ era in genome editing

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