facilitating-cerebral-organoid-culture-via-lateral-soft-light

Facilitating Cerebral Organoid Culture <em>via</em> Lateral Soft Light Illumination

At present, cerebral organoid culture technology is still complicated to operate and difficult to apply on a large scale. It is necessary to find a simple ...

Wuyi University

The Tibet minipig is a rare highland pig breed worldwide and has many applications in biomedical and agricultural research. However, Tibet minipigs are not like domesticated pigs in that their ovulation number is low, which is unfavourable for the collection of zygotes. Partly for this reason, few studies have reported the successful generation of genetically modified Tibet minipigs by zygote injection. To address this issue, we described an efficient way to generate gene-edited Tibet minipigs, the major elements of which include the utilization of synchronized oestrus instead of superovulation to obtain zygotes, optimization of the preparation strategy, and co-injection of clustered regularly interspaced short palindromic repeat sequences associated protein 9 (Cas9) mRNA and single-guide RNAs (sgRNAs) into the cytoplasm of zygotes. We successfully obtained allelic  gene knockout ( ) Tibet minipigs with a typical albino phenotype (i.e., red-coloured eyes with light pink-tinted irises and no pigmentation in the skin and hair) as well as     and     Tibet minipigs with typical phenotypes of albinism and immunodeficiency, which was characterized by thymic atrophy and abnormal immunocyte proportions. The overall gene editing efficiency was 75% for the  single gene knockout, while for  and  dual gene editing, the values were 25% and 75%, respectively. No detectable off-target mutations were observed. By intercrossing F generation minipigs, targeted genetic mutations can also be transmitted to gene-edited minipigs' offspring through germ line transmission. This study is a valuable exploration for the efficient generation of gene-edited Tibet minipigs with medical research value in the future.

Optimization Strategy for Generating Gene-edited Tibet Minipigs by Synchronized Oestrus and Cytoplasmic Microinjection.

In this study, we designed and tested fluorescent probe zy-2 for specific and responsive imaging of pyruvate kinase M2 (PKM2), which can be excited by 419 nm light. A 17-fold enhancement in the responsive emission upon zy-2's binding to PKM2 was observed, imaging of which was successfully recorded in a time- and concentration-dependent manner in PKM2-positive cells. Thus, we obtained a responsive fluorescent probe for the specific and sensitive detection of PKM2, which is innovative in design and applicable to the detection of cancer cells.

Article	Year
Optimization Strategy for Generating Gene-edited Tibet Minipigs by Synchronized Oestrus and Cytoplasmic Microinjection. International journal of biological sciences\| PubMed ID: 31754342	2019
A responsive fluorescent probe for detecting and imaging pyruvate kinase M2 in live cells. Chemical communications (Cambridge, England)\| PubMed ID: 35574810	2022

Yinghui Zhang

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