ABSTRACT
The generation of a tau-V337M point mutation mouse model using gene editing technology can provide an animal model with fast disease progression and more severe symptoms, which facilitate the study of pathogenesis and treatment of Alzheimer's disease (AD). In this study, single guide RNAs (sgRNA) and single-stranded oligonucleotides (ssODN) were designed and synthesized in vitro. The mixture of sgRNA, Cas9 protein and ssODN was microinjected into the zygotes of C57BL/6J mice. After DNA cutting and recombination, the site homologous to human 337 valine (GTG) in exon 11 was mutated into methionine (ATG). In order to improve the efficiency of recombination, a Rad51 protein was added. The female mice mated with the nonvasectomy male mice were used as the surrogates. Subsequently, the 2-cell stage gene edited embryos were transferred into the unilateral oviduct, and the F0 tau-V337M mutation mice were obtained. Higher mutation efficiency could be obtained by adding Rad51 protein. The F0 tau-V337M point mutation mice can pass the mutation on to the F1 generation mice. In conclusion, this study successfully established the first tau-V337M mutation mouse by using Cas9, ssODN and Rad51. These results provide a new method for developing AD mice model which can be used in further research on the pathogenesis and treatment of AD.
Subject(s)
Animals , Male , Female , Mice , Humans , CRISPR-Cas Systems/genetics , RNA, Guide, CRISPR-Cas Systems , Rad51 Recombinase/genetics , Mice, Inbred C57BL , Disease Models, Animal , Recombination, GeneticABSTRACT
OBJECTIVE:To establish a method for the dissolution determination of Tamibarotene tablet. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-water-acetic acid(85∶15∶1,V/V/V)at a flow rate of 1.0 ml/min,detection wavelength was 235 nm,column temperature was 25 ℃,and injection volume was 20 μl. The paddle method was used to determine the dissolution,using pH6.8 phosphate buffer dissolution as medium,medium volume was 900 ml,the rotat-ing speed was 50 r/min,taking samples after 60 min. RESULTS:The linear range of tamibarotene was 0.801-2.804 μg/ml (r=0.999 6);RSDs of precision,stability and reproducibility tests were lower than 3%;recovery was 99.58%-100.90%(RSD=0.25%,n=9);the accumulative dissolution rate of 3 batches of samples was 98.25%,96.54%,92.38%(n=6). CONCLUSIONS:The method simple,accurate with high sensitivity and specificity,and suitable for the dissolution determination of Tamibarotene tablet.
ABSTRACT
Dexmedetomidine( DEX) is a pure potent, highly se-lective and highly specific agonist ofα2-adrenergic receptors with sedative, analgesic and sympatholytic properties. The sedative effect mimics natural sleep of“arousable” and“cooperative” se-dation without respiratory depression. Due to the above properties and advantages, DEX has received adequate attention in clinical practice and its spectrum of application is also expanding. In re-cent years, it is proved that DEX is neuroprotective not only in animal researches but also in clinical studies. The neuroprotec-tion of DEX and its related mechanism will be briefly reviewed in this paper.