Synthetic switch-based baculovirus for transgene expression control and selective killing of hepatocellular carcinoma cells

Lin, Mingyao; Tseng, Yen-Wen; Shen, Chih-Che; Hsu, Mu‐Nung; Hwu, Jih Ru; Chang, C.S.; Yeh, Chung-Ju; Chou, Ming-Yi; Wu, Jaw–Ching; Hu, Yu‐Chen

doi:10.1093/nar/gky447

Cited by 24 publications

(19 citation statements)

References 67 publications

(83 reference statements)

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“…MicroRNAs (miRNAs) are another type of non‐coding RNAs characterized by short length. It can interact with the 3′ untranslated regions (UTR) of the target genes via its microRNA response elements (MREs), which will silence the expression of downstream target genes . The prevalent ceRNA (competing endogenous RNA) hypothesis demonstrates well that lncRNA can competitively interact with miRNA and restore the inhibiting biological function of target gene.…”

Section: Introductionmentioning

confidence: 99%

“…It can interact with the 3′ untranslated regions (UTR) of the target genes via its microRNA response elements (MREs), which will silence the expression of downstream target genes. 12 The prevalent ceRNA (competing endogenous RNA) hypothesis demonstrates well that lncRNA can competitively interact with miRNA and restore the inhibiting biological function of target gene. For instance, LUCAT1 has been found can competitively bind to miR-5582-3p against TCF7L, consequently enhancing Wnt/β-catenin pathway in breast cancer stem cell (BCSCs).…”

Section: Introductionmentioning

confidence: 99%

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LncRNA FGD5‐AS1 can be predicted as therapeutic target in oral cancer

Liu

Yao

Huang

et al. 2020

J Oral Pathology Medicine

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Background Long non‐coding RNAs (lncRNAs) have been extensively studied to participate in the carcinogenesis of various tumors. LncRNA FGD5‐AS1 has been studied as an oncogene in several cancers; however, the role it plays in oral squamous cell carcinoma (OSCC) still remains unclear. Methods Quantitative real‐time polymerase chain reaction (qRT‐PCR) was used to assess relevant RNAs expression. CCK‐8 and colony formation were combined to investigate cell proliferation. Flow cytometry was implemented to test the apoptosis of cell. Wound healing assay and transwell assays were conducted to investigate cell migration and invasion. Western blot assay was conducted to measure relevant protein expression. Results FGD5‐AS1 expression was aberrantly up‐regulated in OSCC tissue and cells. FDG5‐AS1 up‐regulation induced USP21 overexpression advances OSCC development. Knockdown of FGD5‐AS1 inhibited cell growth, migration, and invasion, yet promoted apoptosis. Conclusion FGD5‐AS1 regulates OSCC via competitively binding to miR‐520b against USP21. It could become a potential diagnostic biomarker for OSCC.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

LncRNA FGD5‐AS1 can be predicted as therapeutic target in oral cancer

Liu

Yao

Huang

et al. 2020

J Oral Pathology Medicine

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“…MicroRNAs (miRNAs) are one class of small noncoding RNAs with 17 to 25 nt and can bind to the 3′-untranslated region (3′-UTR) of messenger RNA (mRNAs) to degrade mRNA or inhibit mRNA translation, which implicated in numerous biological processes. 7 Aberrant expression of miRNAs plays important roles in the cancer progression. For instance, miR-409 was found to directly bind to SPIN1 and inhibited human non-small-cell lung cancer progression.…”

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confidence: 99%

hsa_circRNA_100533 regulates GNAS by sponging hsa_miR_933 to prevent oral squamous cell carcinoma

Zhu

Shao

Tang

et al. 2019

J of Cellular Biochemistry

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Oral squamous cell carcinoma (OSCC) is the most common malignant tumor of the head and neck region. Circular RNA (circRNA), as one kind of noncoding RNA, involves in biological processes in diverse cancers. circRNA functions mainly as the microRNA (miRNA) sponge, competitively binding to miRNAs to regulate target gene expressions. However, the expression profiles and roles of circRNAs in OSCC are still unexplored. circRNA microarrays and quantitative real‐time polymerase chain reaction was used to identify the hsa_circRNA_100533 downregulated in OSCC tissues and cell lines. Bioinformatics methods were used to predict the interactions among circRNAs, miRNA, and target genes. Based on the luciferase reporter assay and AGO2 RIP assay, we found that hsa_circRNA_100533 binds to miRNAs as a miRNA sponge. hsa_circRNA_100533 inhibited cell proliferation, migration, and promoted cell apoptosis in OSCC cell lines, which could be blocked by hsa‐miR‐933 overexpression. hsa_circRNA_100533 binds to hsa‐miR‐933 as a miRNA sponge to regulate GNAS expression, and to modulate cell proliferation, migration, and apoptosis. In summary, the hsa_circRNA_100533‐miR‐933‐GNAS axis affect the proliferation and apoptosis of OSCC cells through the mechanism of competing endogenous RNAs. hsa_circRNA_100533 may function as promising diagnostic biomarkers and effective therapeutic targets for OSCC.

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“…Such system allows for the construction of AND, OR, NAND, and NOR dCas9 logic operators (58). With the ability to simultaneously turn ON/OFF genes, our CRISPRai system may be adapted for conditional gene activation/repression and be used to build logic gates to control cell fate or classify cell type (59,60).…”

Section: Discussionmentioning

confidence: 99%

CRISPRai for simultaneous gene activation and inhibition to promote stem cell chondrogenesis and calvarial bone regeneration

Truong

Hsu

Nguyen

et al. 2019

Nucleic Acids Research

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Calvarial bone healing remains difficult but may be improved by stimulating chondrogenesis of implanted stem cells. To simultaneously promote chondrogenesis and repress adipogenesis of stem cells, we built a CRISPRai system that comprised inactive Cas9 (dCas9), two fusion proteins as activation/repression complexes and two single guide RNA (sgRNA) as scaffolds for recruiting activator (sgRNAa) or inhibitor (sgRNAi). By plasmid transfection and co-expression in CHO cells, we validated that dCas9 coordinated with sgRNAa to recruit the activator for mCherry activation and also orchestrated with sgRNAi to recruit the repressor for d2EGFP inhibition, without cross interference. After changing the sgRNA sequence to target endogenous Sox9/PPAR-γ, we packaged the entire CRISPRai system into an all-in-one baculovirus for efficient delivery into rat bone marrow-derived mesenchymal stem cells (rBMSC) and verified simultaneous Sox9 activation and PPAR-γ repression. The activation/inhibition effects were further enhanced/prolonged by using the Cre/loxP-based hybrid baculovirus. The CRISPRai system delivered by the hybrid baculovirus stimulated chondrogenesis and repressed adipogenesis of rBMSC in 2D culture and promoted the formation of engineered cartilage in 3D culture. Importantly, implantation of the rBMSC engineered by the CRISPRai improved calvarial bone healing. This study paves a new avenue to translate the CRISPRai technology to regenerative medicine.

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Synthetic switch-based baculovirus for transgene expression control and selective killing of hepatocellular carcinoma cells

Cited by 24 publications

References 67 publications

LncRNA FGD5‐AS1 can be predicted as therapeutic target in oral cancer

LncRNA FGD5‐AS1 can be predicted as therapeutic target in oral cancer

hsa_circRNA_100533 regulates GNAS by sponging hsa_miR_933 to prevent oral squamous cell carcinoma

CRISPRai for simultaneous gene activation and inhibition to promote stem cell chondrogenesis and calvarial bone regeneration

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