6 results about "Pronucleus" patented technology

The application provides a construction method of a humanized keloid mouse model, comprising the following steps: BAC plasmid construction and preparation; superovulation of experimental mice and collection of zygotes; pronuclear microinjection of zygotes; post-injection embryo transplantation; genotype identification; breeding and genetic analysis; and verification of construction results of the humanized immune system by immunohistochemistry and immunofluorescence. In the application, peripheral blood mononuclear cells treated by sCD27 and skin around keloids (homologous cells and tissues) are transplanted into NSG-MHC-DKO immune-deficient mice with overexpression of CD70 genes, so that HLA rejection of different homologous immune cells and tissues can be avoided, the internal microenvironment of keloids and the interaction between the internal microenvironment and the immune system can be restored to the maximum extent, and the influence of immune factors on the occurrence and development of keloids can be realized in vitro. The application first discovers and verifies that the activation of the CD27-CD70 axis can promote the occurrence and development of keloids, and provides a suitable animal model for the research and development of anti-keloid drugs, especially immunotherapy.

This invention discloses a method and system for embryo quality detection, relating to the field of embryo detection technology, including the following steps: S1: fertilized embryos are continuously cultured in an in vitro culture system containing specific metabolic marker detection components, and continuous dynamic imaging of the embryos is performed to obtain a complete dynamic developmental image sequence; S2: the dynamic developmental image sequence is segmented, and temporal developmental features are captured, identifying key time points such as pronucleus appearance and disappearance, cleavage, and blastocyst formation, and generating a segmentation mask corresponding to each frame of the dynamic developmental image sequence; this invention achieves full-process time-difference imaging to capture the temporal features of embryonic development, without trauma or interference, avoiding subjective bias in static observation; by segmenting the dynamic developmental image sequence and capturing temporal developmental features, the segmentation accuracy is high and the anti-interference ability is strong.

The invention discloses a method for constructing an anti-spinetoram neoseiulus californicus gene and an ovary targeting peptide of the anti-spinetoram neoseiulus californicus gene. According to the invention, a segment of yolk protein targeted peptide binding region is identified in Neoseiulus californicus, and the yolk protein targeted peptide binding region and a Cas9 sequence are fused and constructed on a PET28a carrier, and the NcvgP2C-Cas9 protein is obtained through prokaryotic expression. NcvgP2C-Cas9 protein, sgNcnAChR alpha 6 and an endosome escape reagent are mixed and injected into female adult mites, NcnAChR alpha 6 gene large fragment deleted neoseiulus californicus offspring is obtained and shows a phenotype with 23.2 times of resistance to spinetoram, and the resistance of a knockout strain to the spinetoram belongs to an autosome partial recessive genetic mode. The invention provides a new method for the genetic improvement of the drug resistance of Neoseiulus californicus, and enhances the compatibility between biological control and pesticide treatment strategies.

The application relates to the technical field of hydrogel preparations, and discloses a hydrogel preparation, a preparation method and application in repairing sperm damage. The method comprises the following steps: step one, constructing a recombinant plasmid; step two, plasmid transformation; step three, protein purification; step four, preparing mEXOs wrapped SKAP2 proteins; and step five, preparing a SKAP2 hydrogel preparation. In the application, SKAP2 proteins are expressed by means of a prokaryote Escherichia coli, milk exosomes are extracted, the SKAP2 proteins are then wrapped into the milk exosomes, and the exosomes containing the SKAP2 proteins are then fused into a hydrogel preparation. The sperm activity of asthenospermia mice can be significantly improved through skin smearing, including improving the sperm activity reduction effect of asthenospermia caused by heavy metal lead, organic endocrine disruptors phthalate, high temperature and aging, and the problem that there is no effective repair preparation for asthenospermia in the prior art is solved.

The invention discloses a construction method and application of an IER2-Flag gene knock-in mouse model, and the construction method comprises the following steps: determining a target site of an IER2 gene to be knocked in a mouse, and designing sgRNA with a sequence as shown in SEQ ID No: 2; the method comprises the following steps: carrying out recombination on double-stranded sgRNA formed by annealing and CRISPR-Cas9 plasmid PX459, and constructing a PX459-sgRNA recombinant plasmid of which the sequence is as shown in SEQ ID No: 3-4; the method comprises the following steps: by taking a recombinant plasmid as a template, carrying out PCR (Polymerase Chain Reaction) amplification on a double-stranded DNA (Deoxyribose Nucleic Acid) fragment of a T7 promoter sequence + sgRNA + tracr RNA, and carrying out in-vitro transcription to obtain single-stranded sgRNA + tracr RNA, a single-chain sgRNA + tracr RNA, a Donor sequence containing a 3 * Flag tag and Cas9 Protein are co-injected into a male pronucleus of a mouse fertilized egg, an F0-generation IER2-Flag gene knock-in mouse can be obtained until an F3-generation IER2-Flag gene knock-in mouse is obtained, and the IER2-Flag gene knock-in mouse model is obtained. According to the invention, the expression of the Flag tag is detected for the first time to further identify that the IER2 protein is continuously and strongly expressed in the enamel cell, and an important animal model is provided for the research on revealing a mechanism for regulating and controlling enamel mineralization by the IER2 gene.

Embodiments described herein use deep learning to automate measurement of key morphokinetic embryo features associated with viability and quality, in particular those relevant for clinical in-vitro fertilization (IVF). Systems and methods may, for example, acquire one or more digital images of one or more embryos; select one or more embryos in each digital image; and for each selected embryo, (i) computationally characterize the zona pellucida, detect the degree of fragmentation in the embryo, and for each embryo with a low fragmentation score, computationally classify the embryo's developmental stage based on whether cells constituting the embryo exceed a threshold number (e.g., nine). For an embryo consisting of a single cell, pronuclei may be detected and counted. Based on these measurements, a viability score may be assigned to the embryo.