Hydrodynamics-based liver transfection achieves gene silencing of CB1 using short hairpin RNA plasmid in cirrhotic rats

Díaz-Rivera, Adriana; Meza-Ríos, Alejandra; Sánchez, Victoria Chagoya de; Velasco-Loyden, Gabriela; García-Benavides, Leonel; Jave-Suárez, Luis F; Monroy-Ramírez, Hugo Christian; Santos, Arturo; Armendáriz‐Borunda, Juan; Sandoval-Rodríguez, Ana

doi:10.1371/journal.pone.0228729

Cited by 3 publications

(3 citation statements)

References 30 publications

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“…Animals were anesthetized using ~2% isoflurane in oxygen (UNO BV, UNO, Zevenaar, The Netherlands). Hydrodynamic-based transfection was performed by injecting 100 µg of plasmids into Altogen's liver in vivo transfection reagent (Altogen Biosystems, Las Vegas, NV, USA) via the lateral tail vein [22,23]. The volume of the transfectant mixture was adjusted with 150 mM PBS, pH = 7.2 to 10% of equivalent body weight (1 mL/10 g, 2.5 mL for a 25 g mouse, injected within 7 s).…”

Section: Plasmids and In Vivo Shrna Plasmids Injectionmentioning

confidence: 99%

The Role of Acyl-CoA Synthetase 1 in Bioactive Lipid Accumulation and the Development of Hepatic Insulin Resistance

Zabielski,

Imierska,

Roszczyc-Owsiejczuk

et al. 2024

Nutrients

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The liver plays a crucial role in glucose metabolism. Obesity and a diet rich in fats (HFD) contribute to the accumulation of intracellular lipids. The aim of the study was to explore the involvement of acyl-CoA synthetase 1 (ACSL1) in bioactive lipid accumulation and the induction of liver insulin resistance (InsR) in animals fed an HFD. The experiments were performed on male C57BL/6 mice divided into the following experimental groups: 1. Animals fed a control diet; 2. animals fed HFD; and 3. HFD-fed animals with the hepatic ACSL1 gene silenced through a hydrodynamic gene delivery technique. Long-chain acyl-CoAs, sphingolipids, and diacylglycerols were measured by LC/MS/MS. Glycogen was measured by means of a commercially available kit. The protein expression and phosphorylation state of the insulin pathway was estimated by Western blot. HFD-fed mice developed InsR, manifested as an increase in fasting blood glucose levels (202.5 mg/dL vs. 130.5 mg/dL in the control group) and inhibition of the insulin pathway, which resulted in an increase in the rate of gluconeogenesis (0.420 vs. 0.208 in the control group) and a decrease in the hepatic glycogen content (1.17 μg/mg vs. 2.32 μg/mg in the control group). Hepatic ACSL1 silencing resulted in decreased lipid content and improved insulin sensitivity, accounting for the decreased rate of gluconeogenesis (0.348 vs. 0.420 in HFD(+/+)) and the increased glycogen content (4.3 μg/mg vs. 1.17 μg/mg in HFD(+/+)). The elevation of gluconeogenesis and the decrease in glycogenesis in the hepatic tissue of HFD-fed mice resulted from cellular lipid accumulation. Inhibition of lipid synthesis through silencing ACSL1 alleviated HFD-induced hepatic InsR.

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Section: Plasmids and In Vivo Shrna Plasmids Injectionmentioning

confidence: 99%

The Role of Acyl-CoA Synthetase 1 in Bioactive Lipid Accumulation and the Development of Hepatic Insulin Resistance

Zabielski,

Imierska,

Roszczyc-Owsiejczuk

et al. 2024

Nutrients

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“…Although no protective ingredients are included in its formulation, hydrodynamic delivery can act as an effective carrier, not only of DNA [33,114,124,132,136,137,143,171] but also of other materials, including RNA and proteins, because rapid injection of a large volume of solution physically pushes away preexisting blood components, including nucleases, and brings nucleic acids or proteins directly into the cell interior within a short period of time. Since the establishment of the concept of RNA interference to suppress gene expression, there have been reports of the effective hydrodynamic delivery of various types of RNA, including microRNA [47,150,152,[188][189][190][191], circular RNA [126], short hairpin RNA [94,192,193], and small interfering RNA (siRNA) [87,[194][195][196]. Hydrodynamic delivery can also extrude cells from blood vessels and, thus, offers a simple and easy way to develop animal models for metastatic diseases [118] (Table 3).…”

Section: Delivery Of Materialsmentioning

confidence: 99%

“…Minicircle DNA [33,114,124,132,136,137,143,171] US-targeted microbubble destruction [178] PhiC31 Integrase [197] microRNA [47,150,152,[188][189][190][191] Computer-assisted hydrodynamic delivery [12,198] Sleeping Beauty [30,96,111,142,191,[199][200][201][202] Circular RNA [126] Bioluminescence imaging [47,[203][204][205][206] piggyBac [126,207] shRNA [94,192,193] Tissue clearing [206] Cre-loxP [208][209][210] siRNA [87,[194][195][196] Repopulation [189,211] CreER…”

Section: Delivery Materials Technological Developments Gene Editingmentioning

confidence: 99%

Hydrodynamic Delivery: Characteristics, Applications, and Technological Advances

et al. 2023

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The principle of hydrodynamic delivery was initially used to develop a method for the delivery of plasmids into mouse hepatocytes through tail vein injection and has been expanded for use in the delivery of various biologically active materials to cells in various organs in a variety of animal species through systemic or local injection, resulting in significant advances in new applications and technological development. The development of regional hydrodynamic delivery directly supports successful gene delivery in large animals, including humans. This review summarizes the fundamentals of hydrodynamic delivery and the progress that has been made in its application. Recent progress in this field offers tantalizing prospects for the development of a new generation of technologies for broader application of hydrodynamic delivery.

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Silencing the glycerol-3-phosphate acyltransferase-1 gene in the liver of mice fed a high-fat diet, enhances insulin sensitivity and glucose metabolism by promoting fatty acid beta-oxidation

Zabielski,

Roszczyc-Owsiejczuk,

Imierska

et al. 2024

Biomedicine & Pharmacotherapy

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Hydrodynamics-based liver transfection achieves gene silencing of CB1 using short hairpin RNA plasmid in cirrhotic rats

Cited by 3 publications

References 30 publications

The Role of Acyl-CoA Synthetase 1 in Bioactive Lipid Accumulation and the Development of Hepatic Insulin Resistance

The Role of Acyl-CoA Synthetase 1 in Bioactive Lipid Accumulation and the Development of Hepatic Insulin Resistance

Hydrodynamic Delivery: Characteristics, Applications, and Technological Advances

Silencing the glycerol-3-phosphate acyltransferase-1 gene in the liver of mice fed a high-fat diet, enhances insulin sensitivity and glucose metabolism by promoting fatty acid beta-oxidation

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