genetically modified mice
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PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258942
Author(s):  
Chenyu Zhu ◽  
Rui Xu ◽  
Yuxin Li ◽  
Michael Andrade ◽  
Deng Ping Yin

Obese subjects have increase probabilities of developing type 2 diabetes (T2D). In this study, we sought to determine whether gastric bypass prevents the progression of prediabetes to overt diabetes in genetically modified mice and chemically induced diabetic mice. Roux-en-Y gastric bypass (RYGB) was performed in C57BL/KsJ-db/db null (BKS-db/db,) mice, high-fat diet (HFD)-fed NONcNZO10/LtJ (NZO) mice, C57BL/6 db/db null (B6-db/db) mice and streptozotocin (STZ)-induced diabetic mice. Food consumption, body weight, fat mass, fast blood glucose level, circulating insulin and adiponectin and glucose tolerance test were analyzed. The liver and pancreatic tissues were subjected to H&E and immunohistochemistry staining and islet cells to flow cytometry for apoptotic analysis. RYGB resulted in sustained normoglycemia and improved glucose tolerance in young prediabetic BKS-db/db mice (at the age of 6 weeks with hyperglycemia and normal insulinemia) and HFD-fed NZO and B6-db/db mice. Remarkably, RYGB improved liver steatosis, preserved the pancreatic β-cells and reduced β-cell apoptosis with increases in circulating insulin and adiponectin in young prediabetic BKS-db/db mice. However, RYGB neither reversed hyperglycemia in adult diabetic BKS-db/db mice (12 weeks old) nor attenuated hyperglycemia in STZ-induced diabetic mice. These results demonstrate that gastric bypass improves hyperglycemia in genetically modified prediabetic mice; however, it should be performed prior to β-cells exhaustion.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tomoo Eto ◽  
Hiroki Ueda ◽  
Ryoji Ito ◽  
Tsukasa Takahashi ◽  
Toshiaki Watanabe ◽  
...  

AbstractGenetically modified mice are commonly used in biologic, medical, and drug discovery research, but conventional microinjection methods used for genetic modification require extensive training and practical experience. Here we present a fully automated system for microinjection into the pronucleus to facilitate genetic modification. We first developed software that automatically controls the microinjection system hardware. The software permits automatic rotation of the zygote to move the pronucleus to the injection pipette insertion position. We also developed software that recognizes the pronucleus in 3-dimensional coordinates so that the injection pipette can be automatically inserted into the pronucleus, and achieved a 94% insertion rate by linking the 2 pieces of software. Next, we determined the optimal solution injection conditions (30 hPa, 0.8–2.0 s) by examining the survival rate of injected zygotes. Finally, we produced transgenic (traditional DNA injection and piggyBac Transposon system) and knock-in (genomic editing) mice using our newly developed Integrated Automated Embryo Manipulation System (IAEMS). We propose that the IAEMS will simplify highly reproducible pronuclear stage zygote microinjection procedures.


Author(s):  
Mattia L. DiFrancesco ◽  
Pietro Mesirca ◽  
Isabelle Bidaud ◽  
Dirk Isbrandt ◽  
Matteo E. Mangoni

JCI Insight ◽  
2021 ◽  
Author(s):  
Haixia Zhang ◽  
Alex M. Hanson ◽  
Tobias U. Scherf de Almeida ◽  
Christopher H. Emfinger ◽  
Conor McClenaghan ◽  
...  

3 Biotech ◽  
2021 ◽  
Vol 11 (2) ◽  
Author(s):  
Bita Ghassemi ◽  
Monire Jamalkhah ◽  
Gelareh Shokri ◽  
Mousa Kehtari ◽  
Masoud Soleimani ◽  
...  

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Shun-ichi Funano ◽  
Daisuke Tone ◽  
Hideki Ukai ◽  
Hiroki R. Ueda ◽  
Yo Tanaka

Abstract Objective Production of genetically modified mice including Knock-out (KO) or Knock-in (KI) mice is necessary for organism-level phenotype analysis. Embryonic stem cell (ESC)-based technologies can produce many genetically modified mice with less time without crossing. However, a complicated manual operation is required to increase the number of ESC colonies. Here, the objective of this study was to design and demonstrate a new device to easily find colonies and carry them to microwells. Results We developed a polydimethylsiloxane-based device for easy manipulation and isolation of ESC colonies. By introducing ESC colonies into the groove placed near culturing microwells, users can easily find, pick up and carry ESC colonies to microwells. By hydrophilic treatment using bovine serum albumin, 2-μL droplets including colonies reached the microwell bottom. Operation time using this device was shortened for both beginners (2.3-fold) and experts (1.5-fold) compared to the conventional colony picking operation. Isolated ESC colonies were confirmed to have maintained pluripotency. This device is expected to promote research by shortening the isolation procedure for ESC colonies or other large cells (e.g. eggs or embryos) and shortening training time for beginners as a simple sorter.


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