Synthesizing AND gate genetic circuits based on CRISPR-Cas9 for identification of bladder cancer cells

Liu, Yuchen; Zeng, Yayue; Liu, Li; Zhuang, Cheng‐Le; Fu, Xing; Huang, Weiren; Cai, Zhiming

doi:10.1038/ncomms6393

Cited by 197 publications

(155 citation statements)

References 29 publications

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“…The dCas9 has been used for transcriptional activation and repression [134][135][136][137], for localizing fluorescent protein labels [138], and for recruiting histone modifying enzymes [139,140]. Other applications of Cas9 include building gene circuits [141][142][143], creating new anti-microbials [144], antivirals [145][146][147], and large-scale gain-and loss-of-function screening [148][149][150][151] (Table 1).…”

Section: Genome Editing Applicationsmentioning

confidence: 99%

Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems

Mohanraju

Makarova

Zetsche

et al. 2016

Science

571

460

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Background: Prokaryotes have evolved multiple systems to combat invaders such as viruses and plasmids. Examples of such defence systems include receptor masking, restriction-modification (R-M systems), DNA interference (Argonaute), bacteriophage exclusion (BREX or PGL) and abortive infection, all of which act in an innate, non-specific manner. In addition, prokaryotes have evolved adaptive, heritable immune systems, i.e. clustered regularly interspaced palindromic repeats (CRISPR) and the CRISPR-associated proteins (CRISPR-Cas). Adaptive immunity is conferred by the integration of DNA sequences from an invading element into the CRISPR array (adaptation), which is transcribed into long pre-CRISPR (pre-cr) RNAs and processed into short crRNAs (expression), which guide Cas proteins to specifically degrade the cognate DNA on subsequent exposures (interference). Advances:A plethora of distinct CRISPR-Cas systems are represented in genomes of most archaea and almost half of the bacteria. The latest CRISPR-Cas classification scheme delineates two classes that are each subdivided into three types. Integration of biochemistry and molecular genetics has contributed significantly to revealing many of the unique features of the variant CRISPR-Cas types. Additionally, structural analysis and single molecule studies have further advanced our understanding of the molecular basis of CRISPR-Cas functionality. Recent progress includes relevant steps in the adaptation stage, when fragments of foreign DNA are processed and incorporated as new spacers into the CRISPR array. In addition, three novel CRISPR-Cas types (IV, V, and VI) have been identified, and in particular, the type V interference complexes have been experimentally characterized. Moreover, the ability to easily program sequence-specific DNA targeting and cleavage by CRISPRCas components, as demonstrated for Cas9 and Cpf1, allows for the application of CRISPR-Cas components as highly effective tools for genetic engineering and gene regulation in a wide range of eukaryotes and prokaryotes.The pressing issue of off-target cleavage by the Cas9 nuclease is being actively addressed using structure-guided engineering.Outlook: Although our understanding of the CRISPR-Cas system has increased tremendously over the past few years, much remains to be done. About the evolution of CRISPR-Cas systems, the continuing discovery of novel CRISPR-Cas variants will provide direct tests of the recently proposed modular scenario. The recent discovery and characterization of new CRISPR-Cas types with many unique features implies that our current knowledge has relatively limited power for predicting the functional details of distantly related variants. Hence, newly discovered CRISPR-Cas systems need to be thoroughly dissected with the aforementioned multi-disciplinary approaches to gain insight in their biological role, to unravel their molecular mechanism, and to harness their potential for biotechnology. As to the biology, key outstanding questions include the ecological roles of microbial ...

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Section: Genome Editing Applicationsmentioning

confidence: 99%

Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems

Mohanraju

Makarova

Zetsche

et al. 2016

Science

571

460

View full text Add to dashboard Cite

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“…Zalatan et al reprogrammed a branched metabolic pathway in yeast to control the production of various product metabolites (30). Liu et al used a nuclease Cas9 to create a promoter-based "AND" logic gate to identify and control a specific type of cancer cell (34). Promoter-based logic gates (e.g., AND, OR, and NOT) could also be combined with the scRNA components to create complex regulatory patterns that are induced only under certain conditions.…”

Section: Engineering Complex Regulatory Patterns Using Crispra/imentioning

confidence: 99%

The New State of the Art: Cas9 for Gene Activation and Repression

Russa

2015

Molecular and Cellular Biology

206

150

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CRISPR-Cas9 technology has rapidly changed the landscape for how biologists and bioengineers study and manipulate the genome. Derived from the bacterial adaptive immune system, CRISPR-Cas9 has been coopted and repurposed for a variety of new functions, including the activation or repression of gene expression (termed CRISPRa or CRISPRi, respectively). This represents an exciting alternative to previously used repression or activation technologies such as RNA interference (RNAi) or the use of gene overexpression vectors. We have only just begun exploring the possibilities that CRISPR technology offers for gene regulation and the control of cell identity and behavior. In this review, we describe the recent advances of CRISPR-Cas9 technology for gene regulation and outline advantages and disadvantages of CRISPRa and CRISPRi (CRISPRa/i) relative to alternative technologies.

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“…[7][8][9][10][11][12] However, mutation of lncRNAs will cause cancer initiation and promote the metastasis of malignancy. [13][14][15] For instance, lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was first found in lung cancer metastasis. Many studies had demonstrated that lncRNA MALAT1 was overexpressed in various cancer, and it might act as a potential biomarker and therapeutic target in cancer treatment.…”

Section: Introductionmentioning

confidence: 99%