Synthetic miR-21 decoy circularized by tRNA splicing mechanism inhibited tumorigenesis in glioblastoma in vitro and in vivo models

Bayat, Hadi; Pourgholami, Mohammad H.; Rahmani, S.; Pournajaf, Sanaz; Mowla, Seyed Javad

doi:10.1016/j.omtn.2023.04.001

Cited by 8 publications

(2 citation statements)

References 54 publications

(71 reference statements)

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“…In addition to applications as a biomarker, circRNAs can be constructed to target and sequester overexpressed oncogenic miRNAs. The synthetically generated circRNA CM21D was produced via t-RNA splicing and administered in experimental glioblastoma models, inhibiting miR-21-5p thus restoring the expression of tumor suppressor genes [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

Computational approaches and challenges in the analysis of circRNA data

Digby,

Finn,

Ó Broin

2024

BMC Genomics

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Circular RNAs (circRNA) are a class of non-coding RNA, forming a single-stranded covalently closed loop structure generated via back-splicing. Advancements in sequencing methods and technologies in conjunction with algorithmic developments of bioinformatics tools have enabled researchers to characterise the origin and function of circRNAs, with practical applications as a biomarker of diseases becoming increasingly relevant. Computational methods developed for circRNA analysis are predicated on detecting the chimeric back-splice junction of circRNAs whilst mitigating false-positive sequencing artefacts. In this review, we discuss in detail the computational strategies developed for circRNA identification, highlighting a selection of tool strengths, weaknesses and assumptions. In addition to circRNA identification tools, we describe methods for characterising the role of circRNAs within the competing endogenous RNA (ceRNA) network, their interactions with RNA-binding proteins, and publicly available databases for rich circRNA annotation.

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Section: Introductionmentioning

confidence: 99%

Computational approaches and challenges in the analysis of circRNA data

Digby,

Finn,

Ó Broin

2024

BMC Genomics

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“…Consequently, miRNAs are regarded as particularly desirable biomarkers for cancer diagnosis and prognosis. 18,19 Traditionally, northern blotting serves as the gold standard method for miRNA analysis, but suffers from radiolabeling and complicated operation steps. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) approach is widely used for miRNA detection owing to its high sensitivity; however, it requires specialized equipment, professional operation, and prolonged assay time.…”

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confidence: 99%

A lateral flow assay strip for simultaneous detection of miRNA and exosomes in liver cancer

Wei,

Wang,

Zhou

et al. 2024

Chem. Commun.

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CHO cell engineering via targeted integration of circular miR-21 decoy using CRISPR/RMCE hybrid system

Adibzadeh,

Amiri,

Barkhordari

et al. 2024

Appl Microbiol Biotechnol

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Chinese hamster ovary (CHO) cells, widely acknowledged as the preferred host system for industrial recombinant protein manufacturing, play a crucial role in developing pharmaceuticals, including anticancer therapeutics. Nevertheless, mammalian cell-based biopharmaceutical production methods are still beset by cellular constraints such as limited growth and poor productivity. MicroRNA-21 (miR-21) has a major impact on a variety of malignancies, including glioblastoma multiforme (GBM). However, reduced productivity and growth rate have been linked to miR-21 overexpression in CHO cells. The current study aimed to engineer a recombinant CHO (rCHO) cell using the CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) system coupled with the Bxb1 recombinase-mediated cassette exchange (RMCE) to express a circular miR-21 decoy (CM21D) with five bulged binding sites for miR-21 sponging. Implementing the ribonucleoprotein (RNP) delivery method, a landing pad was inserted into the genome utilizing the CRIS-PITCh technique. Subsequently, the CM21D cassette flanked by Bxb1 attB was then retargeted into the integrated landing pad using the RMCE/Bxb1 system. This strategy raised the targeting efficiency by 1.7-fold, and off-target effects were decreased. The miR-21 target genes (Pdcd4 and Atp11b) noticed a significant increase in expression upon the miR-21 sponging through CM21D. Following the expression of CM21D, rCHO cells showed a substantial decrease in doubling time and a 1.3-fold increase in growth rate. Further analysis showed an increased yield of hrsACE2, a secretory recombinant protein, by 2.06-fold. Hence, we can conclude that sponging-induced inhibition of miR-21 may lead to a growth rate increase that could be linked to increased CHO cell productivity. For industrial cell lines, including CHO cells, an increase in productivity is crucial. The results of our research indicate that CM21D is an auspicious CHO engineering approach. Key points • CHO is an ideal host cell line for producing industrial therapeutics manufacturing, and miR-21 is downregulated in CHO cells, which produce recombinant proteins. • The miR-21 target genes noticed a significant increase in expression upon the miR-21 sponging through CM21D. Additionally, sponging of miR-21 by CM21D enhanced the growth rate of CHO cells. • Productivity and growth rate were increased in CHO cells expressing recombinant hrs-ACE2 protein after CM21D knocking in. Graphical abstract

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Synthetic miR-21 decoy circularized by tRNA splicing mechanism inhibited tumorigenesis in glioblastoma in vitro and in vivo models

Cited by 8 publications

References 54 publications

Computational approaches and challenges in the analysis of circRNA data

Computational approaches and challenges in the analysis of circRNA data

A lateral flow assay strip for simultaneous detection of miRNA and exosomes in liver cancer

CHO cell engineering via targeted integration of circular miR-21 decoy using CRISPR/RMCE hybrid system

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