Overhang molecular beacons encapsulated in tethered cationic lipoplex nanoparticles for detection of single-point mutation in extracellular vesicle-associated RNAs

Hu, Jiaming; Kwak, Kwang Joo; Shi, Junfeng; Yu, Bohao; Yan, Sheng; Lee, Ly James

doi:10.1016/j.biomaterials.2018.08.038

Cited by 22 publications

(18 citation statements)

References 30 publications

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“…It has been suggested that tethered cationic lipoplex nanoparticle (tCLN) biochip combined with total internal reflection fluorescence (TRIF) microscopy is an approach which can specifically quantify RNA in exosomes . Therefore, in this study, this method had been employed to demonstrate whether gRNA was present in the cell‐derived exosomes.…”

Section: Resultsmentioning

confidence: 99%

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Friend or Foe? Evidence Indicates Endogenous Exosomes Can Deliver Functional gRNA and Cas9 Protein

Chen

Huang

Liu

et al. 2019

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The clustered regularly interspaced short palindromic repeats (CRISPR)/associated nuclease (Cas) system is an efficient gene editing tool. In this study, it is found that both single guide RNA (gRNA) and Cas9 protein could be exported from the CRISPR/Cas9‐expressing cells by endogenous exosomes independently. Further experiments demonstrate that these naturally produced endogenous exosomes could be used as a vehicle to deliver the functional Cas9 and hepatitis B virus (HBV)‐specific gRNA to cut HBV DNA transfected in HuH7 cells or human papilloma virus (HPV)‐specific gRNA to cut the integrated HPV DNA in HeLa cells, respectively. In conclusion, this study indicates the potential of endogenous exosomes as a safe and effective delivery vehicle of the functional gRNA and Cas9 protein. Meanwhile, the endogenous exosomes‐mediated delivery of gene editing activity to adjacent and distant cells or tissues may further complicate the off‐target and safety concerns about the CRISPR/Cas9 system.

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Section: Resultsmentioning

confidence: 99%

“…tCLN : The detection of exosomal RNA using tCLNs was previously described . Briefly, to make tCLN biochips, a thin Au layer (15 nm) was deposited on a glass cover slip using a Denton e‐beam evaporator (DV‐502A, Denton Vacuum, NJ).…”

Section: Methodsmentioning

confidence: 99%

Friend or Foe? Evidence Indicates Endogenous Exosomes Can Deliver Functional gRNA and Cas9 Protein

Chen

Huang

Liu

et al. 2019

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“…When serum is applied to the biochip, the molecular beacons recognize target miRs in exosomes, and the interaction is read by a total internal reflection fluorescence microscope (15). The copy numbers of the exmiRs can then be obtained by the fluorescence signals of the molecular beacons (MBs) (15). Wu et al (13) demonstrated that the levels of exmiRs detected by the TCLN biochip were consistent with the widely used reverse transcription-quantitative PCR (RT-qPCR) method.…”

Section: Introductionmentioning

confidence: 94%

“…Molecular beacons in liposome nanoparticles serve as molecular probes for target miRs. When serum is applied to the biochip, the molecular beacons recognize target miRs in exosomes, and the interaction is read by a total internal reflection fluorescence microscope (15). The copy numbers of the exmiRs can then be obtained by the fluorescence signals of the molecular beacons (MBs) (15).…”

Section: Introductionmentioning

confidence: 99%

Elevated expression of exosomal microRNA‑21 as a potential biomarker for the early diagnosis of pancreatic cancer using a tethered cationic lipoplex nanoparticle biochip

Ding

et al. 2020

Oncol Lett

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Pancreatic cancer (PC) has a poor prognosis due to the lack of effective molecular biomarkers for early diagnosis. Recent studies have investigated the use of exosomal microRNAs (exmiRs) as diagnostic biomarkers in cancer. The present study examined exmiR-21, exmiR-10b and exmiR-212-3p expression in patients with PC and healthy individuals. The expression levels of exmiR-21, exmiR-10b and exmiR-212-3p were examined in the peripheral blood plasma of 36 patients with PC and 65 healthy controls, using tethered cationic lipoplex nanoparticle biochip. The levels of exmiR-21 in the plasma of 34 mice were also evaluated. The expression levels of exmiR-21 and exmiR-10b were significantly greater in patients with PC compared with the control group. The receiver operating characteristic (ROC) analysis indicated that exmiR-21 had better diagnostic performance (P=0.0003; AUC, 0.7171) compared with the other two exmiRs. The diagnostic value of exmiR-21 improved when combined with exmiR-10b (P<0.0001; AUC, 0.791). Furthermore, exmiR-21 was capable of distinguishing patients with early-stage PC from controls and advanced-stage PC (P<0.05, early stage vs. healthy; P<0.001, early stage vs. advanced stage). The results of the present study revealed that the plasma levels of exmiR-21 and exmiR-10b were upregulated in patients with PC. The ROC analyses indicated that exmiR-21 had the best diagnostic performance among the three exmiRs. Furthermore, exmiR-21 was capable of discriminating patients with early-stage PC from healthy controls. These findings indicate the potential of determining the expression of exmiR-21 from serum using a tethered cationic lipoplex nanoparticle biochip as a novel non-invasive strategy for the early diagnosis of PC.

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“…Zinc transporter ZIP4 [ 37 ], cytoskeleton-associated protein 4, a novel Dickkopf1 receptor [ 96 ], and annexin A1 [ 200 ] are also potential biomarkers. A panel of miRNAs, mRNAs, and proteins derived from EVs is useful for the early diagnosis of pancreatic cancer [ 201 , 202 , 203 , 204 , 205 , 206 , 207 ]. It may be possible to predict treatment outcomes using EVs [ 208 ].…”

Section: Potential Of Evs For Liquid Biopsymentioning

confidence: 99%

Anti-Cancer Role and Therapeutic Potential of Extracellular Vesicles

Tominaga

2021

Cancers

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Cell–cell communication is an important mechanism in biological processes. Extracellular vesicles (EVs), also referred to as exosomes, microvesicles, and prostasomes, are microvesicles secreted by a variety of cells. EVs are nanometer-scale vesicles composed of a lipid bilayer and contain biological functional molecules, such as microRNAs (miRNAs), mRNAs, and proteins. In this review, “EVs” is used as a comprehensive term for vesicles that are secreted from cells. EV research has been developing over the last four decades. Many studies have suggested that EVs play a crucial role in cell–cell communication. Importantly, EVs contribute to cancer malignancy mechanisms such as carcinogenesis, proliferation, angiogenesis, metastasis, and escape from the immune system. EVs derived from cancer cells and their microenvironments are diverse, change in nature depending on the condition. As EVs are thought to be secreted into body fluids, they have the potential to serve as diagnostic markers for liquid biopsy. In addition, cells can encapsulate functional molecules in EVs. Hence, the characteristics of EVs make them suitable for use in drug delivery systems and novel cancer treatments. In this review, the potential of EVs as anti-cancer therapeutics is discussed.

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Overhang molecular beacons encapsulated in tethered cationic lipoplex nanoparticles for detection of single-point mutation in extracellular vesicle-associated RNAs

Cited by 22 publications

References 30 publications

Friend or Foe? Evidence Indicates Endogenous Exosomes Can Deliver Functional gRNA and Cas9 Protein

Friend or Foe? Evidence Indicates Endogenous Exosomes Can Deliver Functional gRNA and Cas9 Protein

Elevated expression of exosomal microRNA‑21 as a potential biomarker for the early diagnosis of pancreatic cancer using a tethered cationic lipoplex nanoparticle biochip

Anti-Cancer Role and Therapeutic Potential of Extracellular Vesicles

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