33 results about "Wild Type Mouse" patented technology

Developing vehicles that efficiently deliver genes throughout the human central nervous system (CNS) will broaden the range of treatable genetic diseases. Applicants engineered an AAV capsid, BI-hTFRl, that binds human Transferrin Receptor (TfRl), a protein expressed on the blood-brain barrier (BBB). BI-hTFRl was actively transported across a human brain endothelial cell layer and, relative to AAV9, provided 40-50 times greater reporter expression in the CNS of human TFRC knock-in mice. The enhanced tropism was CNS-specific and absent in wild type mice. When used to deliver GBA1, mutations of which cause Gaucher disease and are linked to Parkinson's disease, BI-hTFRl substantially increased brain and cerebrospinal fluid glucocerebrosidase activity compared to AAV9.

The invention discloses a construction method of a COL4A5-K229X point mutation X-linked Alport syndrome mouse model. The construction method comprises the following steps: aiming at c.685Agt of a No.12 exon of a mouse COL4A5 gene; carrying out T point mutation, and designing and preparing Cas9 mRNA, gRNA and a donor vector; the components are mixed and then microinjected into fertilized eggs of a C57BL / 6J mouse to obtain an F0-generation mouse; identifying the genotype through PCR (Polymerase Chain Reaction) amplification and Sanger sequencing, and screening positive mice; mating the positive F0-generation mice with the wild-type mice, and breeding F1-generation and subsequent generations; the phenotype of the model is further verified through qPCR, biochemical analysis, light microscopic examination, transmission electron microscope and immunofluorescence. The model constructed by the invention shows hematuria, proteinuria, azemia, podocyte loss and irregular thickening and layering of glomerular basement membrane, is consistent with phenotypes of human XLAS patients, and provides an animal model tool for analyzing pathogenesis and developing treatment strategies.

The application discloses application of TESC as an Alzheimer's disease prevention and treatment target and relates to the field of biological medicines. A TESC overexpression (TESC-OE) hippocampal neuron cell line is constructed through lentivirus transfection, and the result shows that TESC-OE has the ability to resist A beta-induced neuron apoptosis and synapse damage. A TESC overexpression model in the hippocampus of a mouse is constructed through stereotactic injection of an adeno-associated virus, and the result shows that the hippocampal gray matter volume in the brain of the TESC-OE mouse is significantly larger than that of a wild type mouse. A beta stereotactic modeling is performed on the basis of TESC overexpression in the hippocampus, and the result shows that the TESC-OE mouse can significantly resist A beta-induced hippocampal atrophy, learning and memory dysfunction and impaired synaptic plasticity. Immunoblotting analysis reveals the molecular mechanism that TESC can increase the expression of anti-apoptotic genes in the hippocampus and reduce the expression of pro-apoptotic genes to exert a neuroprotective effect.

The application provides a method for constructing a Bmp4 gene knock-in mouse model, and the method comprises the following steps: obtaining sgRNA12 shown in SEQ ID NO. 3; mixing the sgRNA12, Cas9 mRNA and a targeting vector shown in SEQ ID NO. 1, and then microinjecting into a mouse zygote to obtain F0 generation mice; selecting F0 generation positive mice in genotype identification results of the F0 generation mice, and mating the F0 generation positive mice with wild type mice to obtain F1 generation mice with stable genotypes; and screening gene targeting mice with correct recombination, and mating the gene targeting mice with retinal specific knock-out tool mice Six3-Cre to obtain Bmp4 gene knock-in model mice. The method has high gene editing efficiency, and the transgenic mouse can highly express human Bmp4 protein.

A mouse model with conditional knockout of a vitamin D-binding protein gene, and the use thereof. A method comprises the following steps: S1, performing flox modification on the mouse vitamin D-binding protein gene to obtain F0-generation positive mice having undergone the flox modification; S2, mating said F0-generation positive mice with wild type mice to obtain F1-generation positive mice with preserved flox modification; S3, hybridizing the F1-generation positive mice with tissue-specific Cre tool mice to obtain heterozygous mice, and mutually mating the heterozygous mice to obtain homozygous mice; and S4, selecting the obtained homozygous mice to undergo tamoxifen injection, so as to obtain mice with conditional knockout of the vitamin D-binding protein gene.

The invention discloses a construction method and application of a DNAJB6a gene specific knockout mouse model, and belongs to the technical field of gene engineering. Comprising the following steps: S1, designing gRNA sequences aiming at ninth to tenth exon regions of the mouse DNAJB6 gene; s2, in vitro transcription is carried out to prepare Cas9mRNA and gRNA; s3, carrying out microinjection on Cas9mRNA and gRNA (guide Ribonucleic Acid) into a fertilized egg of the mouse; s4, transplanting the fertilized ovum after injection into the body of a pseudo-pregnant female mouse to obtain an F0-generation mouse; s5, screening positive F0-generation mice through genotype identification, and mating the positive F0-generation mice with the wild type mice to obtain F1-generation heterozygote mice; s6, the F1-generation heterozygote mice are matched to obtain F2-generation homozygote mice, and the DNAJB6a gene specific knockout mouse model is established. The construction method for constructing the DNAJB6a gene knockout mouse model, provided by the invention, is simple and convenient to operate, short in construction process time and stable in genotype, and a DNAJB6a gene knockout mouse can be effectively obtained.

The invention provides a premature senility mouse and application thereof. The invention also provides a construction method of the premature senility mouse. Compared with a wild type mouse, the Lmna gene of the premature senility mouse has A604V mutation. The invention also provides a nucleic acid molecule for coding the Lmna. According to the nucleic acid molecule, C at the 1811th site is mutated into T, and C at the 1827th site is mutated into T, corresponding to the nucleotide sequence as shown in SEQ ID NO: 1. The premature senility mouse constructed by the method has senility phenotype about 3 months after birth, the service life can reach about 5-12 months, the window phase for research is prolonged, and the practicability of the premature senility mouse is further improved.

The application relates to a construction method of a conditional overexpression ZFP595 transgenic mouse model and application thereof, and relates to the technical field of transgenesis.The construction method comprises the following steps: constructing a homologous recombination vector containing a ZFP595 gene, wherein the vector comprises a 3.3 kb 5' homologous arm, CAG-LSL-Zfp595-HA-IRES-tdTomato-Wpre-pA and a 3.3 kb 3' homologous arm; microinjecting Cas9 mRNA, gRNA and the homologous recombination vector into a zygote of a recipient mouse to obtain F0 generation mice, and performing identification; mating the F0 generation mice identified as positive with wild type mice and backcrossing to obtain a mouse model for conditionally overexpressing ZFP595.The conditional overexpression ZFP595 mouse model constructed by the construction method can be crossed with various Cre mice to realize specific overexpression in different tissues, organs and cells.

The invention belongs to the technical field of biological medicines, and relates to application of Kir4.2 in treatment of renal fibrosis. The invention discovers that the expression level of the Kir4.2 protein (formed by KCNJ15 gene coding) in renal tissues of a fibrotic mouse is obviously lower than that of a wild type mouse for the first time, and the difference has statistical significance; moreover, after the Kir4.2 is specifically knocked out of the renal proximal tubule, renal fibrosis under a UUO and FA induction model can be aggravated, and the fibrosis process of human renal tubule epithelial cells induced by TGF-beta1 in vitro can be remarkably reversed by overexpression of the Kir4.2. Therefore, Kir4.2 can be used as a drug, a drug target or a target gene in gene therapy, is applied to prevention, alleviation and treatment of renal fibrosis, and can provide a new strategy for prevention and treatment of renal fibrosis. The invention further provides a construction method of the kidney proximal tubule specificity Kir4.2 knockout animal model, and the constructed animal model can be used for researching renal fibrosis pathogenesis and screening drugs for preventing, relieving or / and treating renal fibrosis and has a wide application prospect.

The application relates to the technical field of animal model construction, and particularly discloses a Sypl1 gene knockout colorectal cancer mouse model construction method and application, which comprises the following steps: synthesizing a specific target site gRNA of a Sypl1 gene, mixing Cas9 protein and the target site gRNA to obtain an injection compound, microinjecting the injection compound into mouse zygotes, and transplanting the surviving zygotes into the oviducts of pseudopregnant female mice to obtain F0 generation mice, and the mice born after 20 days are Sypl1 gene knockout mouse animal models; and a chemical induction modeling method is used to construct a colorectal cancer model in wild type mice and gene knockout mice. The Sypl1 gene knockout mouse is constructed by using a CRISPR / Cas mediated genome engineering technology Sypl1 combined with the most widely used colorectal cancer chemical inducer azoxymethane / dextran sodium sulfate, a new colorectal cancer mouse model is constructed, compared with a traditional chemical induction model, the tumor formation rate can be significantly improved, and the tumor formation time can be shortened, and a more efficient animal model is provided for colorectal cancer research.

The invention discloses an application of PDGF-C (Platelet Derived Growth Factor-C) in muscle injury repair. Research finds that in a CTX model mouse with Pdgf-c knocked out, the number of proliferated muscle satellite cells (Pax7 + Ki67 +) is remarkably smaller than that of wild type mice. Through AAV-mediated PDGF-C overexpression, the number of CTX model mouse muscle stem cells can be significantly increased, the activation level of the muscle stem cells is improved, and the self-renewal ability of the muscle stem cells is enhanced. In addition, PDGF-C overexpression also significantly increases the area of muscle fibers of CTX model mice and improves the regeneration and repair ability of the muscle fibers, thereby promoting the healing of muscle injury and having a therapeutic effect on the disease. In addition, the effect of the PDGF-C on promoting muscle injury repair is obviously higher than that of the PDGF-B. Therefore, the invention provides a new method for the treatment of muscle injury, and a new way is possibly opened up for the treatment of other muscle-related diseases.

The invention discloses a construction method and application of a Clock gene site-directed mutagenesis mouse model, and belongs to the technical field of basic medical animal models. The construction method comprises the following steps: designing and constructing a gRNA vector, synthesizing a Donor Oligo sequence, and co-injecting the gRNA vector, a Cas9 vector and the synthesized Donor Oligo into a fertilized egg to obtain an F0-generation transgenic mouse; and mating the F0-generation mouse with the wild-type mouse to obtain an F1-generation transgenic mouse. Phenotype analysis on F1-generation transgenic mice finds that Clock gene site-directed mutagenesis mice show age-progressing heart failure phenotypes which are basically consistent with heart failure caused by aging in a natural physiological state, so that the mice can be used as a model for chronic heart failure diseases, and the pathogenic mechanism of the mice is expected to be deeply researched on the basis of the model; therefore, technical support is provided for prevention and treatment of the disease.

The application discloses a construction method and application of a Clock gene site-directed mutant mouse model, and belongs to the technical field of basic medical animal models. The construction method comprises the following steps: gRNA vector design and construction, synthesis of a Donor Oligo sequence, co-injection of the gRNA vector, a Cas9 vector and the synthesized Donor Oligo into a fertilized egg to obtain F0 generation transgenic mice; and crossbreeding of the F0 generation mice with wild type mice to obtain F1 generation transgenic mice. Phenotype analysis on the F1 generation transgenic mice shows that the Clock gene site-directed mutant mouse exhibits an age progression heart failure phenotype, which is basically consistent with heart failure caused by aging in a natural physiological state, and therefore, the mouse can be used as a model of chronic heart failure disease, and is expected to be used for in-depth research on the pathogenesis based on the model, thereby providing technical support for prevention and treatment of the disease.

The invention relates to the technical field of animal model construction, and particularly discloses a construction method and application of a Sypl1 gene knockout colorectal cancer mouse model, and the construction method comprises the following steps: synthesizing a specific target site gRNA of a Sypl1 gene, uniformly mixing Cas9 protein and the target site gRNA to obtain an injection compound, microinjecting the injection compound into a mouse fertilized egg to obtain a Sypl1 gene knockout colorectal cancer mouse model. And transplanting the survival fertilized ova into the salpingtube of a pseudopregnant female mouse for 20 days to obtain a mouse which is an F0-generation mouse, obtaining a Sypl1 gene knockout mouse animal model, and constructing a colorectal cancer model in a wild type mouse and a gene knockout mouse by adopting a chemical induction modeling method. A Sypl1 gene knockout mouse is constructed through a CRISPR / Cas mediated genome engineering technology, a novel colorectal cancer mouse model is constructed in combination with a colorectal cancer chemical inducer azomethane / dextran sodium sulfate which is most widely applied at present, and compared with a traditional chemical induction model, the novel colorectal cancer mouse model has the advantages that the tumor formation rate can be remarkably increased, the tumor formation time can be shortened, and the tumor formation cost can be reduced. And a more efficient animal model is provided for colorectal cancer research.

The invention discloses an application of fetal calf serum exosome in preparation of a medicine for treating UV irradiation skin injury. When the high-activity exosome derived from the fetal calf serum is used for performing intervention treatment on a C57 wild type mouse UV irradiation skin injury model, the result shows that the exosome derived from the fetal calf serum can remarkably promote pathological improvement of the UV irradiation skin injury model and promote repair of ulceration caused by skin ultraviolet radiation; the regeneration of the epidermal layer and the corium layer of the skin is promoted. The effect of improving and treating the skin injury caused by UV irradiation is achieved, and no obvious side effect exists. Generally speaking, the obtained fetal calf serum-derived high-activity exosome can be used as an effective means for treating UV irradiation skin injury.

The invention belongs to the technical field of biology, and particularly relates to a construction method of a mouse model with hyperuricemia accompanied by hyperglycemia and liver function impairment. The method comprises the following steps: designing double sgRNAs according to the front side and the rear side of a Uox gene Exon 2-4, uniformly mixing the sgRNAs required by gene knockout with Cas9mRNA, injecting the mixture into a fertilized egg of a mouse through microinjection, culturing for 2-4 hours, and transplanting an embryo into a pregnant female mouse to finally obtain an F0 generation mouse. PCR identification and sequencing analysis are carried out, proper Uox gene positive knockout mice are screened to be combined with wild type mice to obtain an F1 generation, and then heterozygous female mice in the F1 generation are selected to mate with heterozygous male mice to obtain homozygous F2 generation mice. And finally determining that the mouse model with hyperuricemia accompanied by hyperglycemia and liver function impairment is successfully constructed through identification.

A construction method of a low-modification psoriasis mouse model based on dihydroxyisobutyrylation of ACTR3 42-site lysine comprises the following steps: step 1, obtaining Cas9mRNA and guideRNA in an in-vitro transcription mode, and obtaining oligo donor DNA through synthesis; step 2, carrying out micro-injection on the Cas9mRNA, guideRNA and oligo donor DNA obtained in the step 1 into a fertilized egg male pronucleus of a C57BL / 6J mouse, transplanting the fertilized egg after injection into the uterus of a pseudopregnant female mouse, taking the mouse born for about 20 days as an F0-generation mouse, and carrying out genotype identification on the F0-generation mouse through PCR (Polymerase Chain Reaction) amplification and sequencing; thirdly, the identified F0-generation positive mice are mated with C57BL / 6J mice, F1-generation mice are obtained through breeding, genotype identification is conducted on the F1-generation mice through PCR amplification and sequencing, and the F1-generation positive mice can be stably inherited heterozygous mice with ACTR3K42A point mutation; the F1-generation negative mouse can be used as a wild type mouse; and 4, carrying out imiquimod induction on the ACTR3K42A point mutation heterozygote mouse obtained in the step 3 to obtain the psoriasis mouse model.

The invention discloses application of alpha1-acid glycoprotein in preparation of a medicine for treating lung injury caused by acute pancreatitis, and belongs to the technical field of biological medicine. An acute pancreatitis lung injury model is constructed through ORM1 gene knockout mice, the mice are divided into a normal group, a model group and a treatment group for research, target organ pathological injury degree detection, immunohistochemical analysis and serum inflammatory factor expression quantity are performed after injection administration, and the curative effect difference of the gene knockout mice and wild type mice is compared, so that the acute pancreatitis lung injury model is established. And the treatment effect of the alpha1-acid glycoprotein on the acute pancreatitis lung injury is verified. The invention provides a key basis for research, development and preparation of drugs for treating acute pancreatitis lung injury, and has important clinical application value.

The invention discloses a construction method and application of a TET2 gene knockout MDS model mouse, and the method is as follows: gRNA and Cas9mRNA of a targeted TET2 gene knockout region, and second gRNA and donor DNA fragments of a targeted flox site are microinjected into a mouse fertilized egg together, the fertilized egg is transplanted into a pseudopregnant female mouse body, and flox positive F0 generation mouse is obtained after farrowing; the F0-generation mice and the wild type mice are subjected to mating to obtain F1-generation Tet2-flox hybrid mice, and the F1-generation Tet2-flox hybrid mice and the wild type mice are subjected to mating to obtain Tet2-flox homozygous mice; the Tet2-flox heterozygous mouse is matched with an Mx1-cre positive mouse to obtain a Tet2-flox heterozygous Mx1-cre positive mouse, and the Tet2-flox heterozygous Mx1-cre positive mouse is matched with a Tet2-flox homozygous mouse to obtain an MDS model mouse with the TET2 gene knocked out. The method is simple in model construction, high in model forming rate and easy to operate.

The invention provides a construction method and application of a dystocia mouse model, and belongs to the technical field of gene engineering. Based on the CRISPR Cas9 technology, gRNA1 with the sequence shown as SEQ ID No.2 and gRNA2 with the sequence shown as SEQ ID No.3 are designed at first, then mRNA of gRNA1, gRNA2 and Cas9 nuclease is injected into a mouse fertilized egg to obtain an F0-generation mouse, then the F0-generation mouse and a wild type mouse are subjected to mating identification and screening to obtain an F1-generation heterozygote mouse, the F1-generation heterozygote female mouse and an F1-generation heterozygote male mouse are subjected to mating identification and screening to obtain a homozygote mouse, and the homozygote mouse and the wild type mouse are subjected to copulation identification and screening to obtain the gRNA1, gRNA2 and Cas9 nuclease. The construction method of the mouse model is simple, the success rate is high, dystocia phenotypes appear after the mouse model is mated, and a reliable animal model is provided for subsequent research on a dystocia triggering mechanism and dystocia treatment.

The invention discloses a congenital cataract mouse model with RagA GTPase point mutation, a construction method and application, and belongs to the technical field of gene editing. The method comprises the following steps: aiming at a gRNA sequence of a mouse RagA GTPase gene, jointly introducing gRNA and Cas9 mRNA into a mouse fertilized egg, and transplanting the fertilized egg into the uterus of a pseudopregnant female mouse to obtain an F0-generation mouse carrying RagA GTPase point mutation; and hybridizing the F0 generation mouse with a wild type mouse, and screening to obtain an F1 generation homozygote mouse carrying RagA GTPase point mutation. According to the invention, a RagA GTPase Leu60Arg point mutation mouse model is constructed by using a CRISPR-Cas9 technology for the first time, and the RagA GTPase Leu60Arg point mutation mouse model can simulate clinical phenotypic characteristics of human congenital cataract and is used for pathogenesis research.

Mammals typically heal with fibrotic scars. Treatments to regenerate human skin and hair without a scar remain elusive. Mice lacking C-X-C motif chemokine receptor 2 (CXCR2-KO) displayed robust and complete tissue regeneration across three different injury models, including skin, hair follicle, and cartilage. Remarkably, wild type mice receiving plasma from CXCR2-KO mice through parabiosis or injections healed wounds scarlessly. A comparison of circulating proteins using multiplex ELISA revealed a 24-fold higher plasma level of granulocyte-colony stimulation factor (G-CSF) in CXCR2-KO blood. Local injections of G-CSF into WT mouse wound beds reduced scar formation and increased hair follicle regeneration by 6-fold. G-CSF directly polarized macrophages into an anti-inflammatory phenotype, and both CXCR2-KO and G-CSF-treated mice recruited more anti-inflammatory macrophages into injured areas. These results improve our molecular understanding of scarless tissue regeneration and introduce a new therapeutic approach for cutaneous wounds and hair regeneration. Provided are compositions and methods relating to treating wound healing pathologies.

The invention discloses an application of PDGF-C in corneal injury repair. Research finds that in a corneal injury model mouse with Pdgf-c knockout, the corneal healing area in the corneal injury mouse model is reduced through Pdgf-c knockout, and the number of corneal limbal stem cells with proliferative activity is remarkably smaller than that of wild type mice. Through AAV-mediated PDGF-C overexpression, proliferation of corneal limbal stem cells can be significantly promoted, and the corneal healing area in a corneal injury mouse model is significantly increased. The results show that overexpression of the PDGF-C significantly improves the healing ability to corneal injury. Therefore, the invention provides a novel method for treating corneal injury.

The application provides a premature aging mouse and application thereof. The application also provides a construction method of the premature aging mouse. Compared with a wild type mouse, the Lmna gene of the premature aging mouse has a mutation of A604V. The application also provides a nucleic acid molecule encoding Lmna, wherein the nucleic acid molecule corresponds to a mutation of C to T at position 1811 and a mutation of C to T at position 1827 in the nucleotide sequence shown in SEQ ID NO:1. The premature aging mouse constructed by the application has an aging phenotype at about 3 months after birth, and the life span can reach about 5-12 months, the window period for research is prolonged, and the practicability of the premature aging mouse is further increased.

The invention belongs to the technical field of biology, and particularly relates to a mouse model for systemic expression or tissue specific expression of KRAS-G12D-EGFP fusion protein and a preparation method thereof, and the preparation method comprises the following steps: a) screening gRNA target sites; b) constructing a homologous recombinant vector; c) mixing the homologous recombinant vector, gRNA and Cas9 mRNA, injecting the mixture into a mouse fertilized egg, and transplanting the mixture into a mouse blastocyst; d) carrying out PCR (Polymerase Chain Reaction) screening on the F0-generation mice to obtain correct homologous recombination positive mice; and e) mating the positive F0-generation mouse with a wild type mouse, breeding, and carrying out PCR screening to obtain an F1-generation recombinant positive mouse. A mouse model modified by the gene is stable in heredity and can be used for researching the biological function of the KRAS-G12D mutant protein in a mouse body. Particularly, the mouse model can be used for researching the function of the KRAS-G12D mutant protein at the tissue specificity level, and the mutant protein can be subjected to cell tissue tracing by using EGFP fluorescent protein, so that the mouse model has a certain application value.

The invention relates to the field of biomedicine, in particular to a disease model construction method based on intestinal epithelium specific knockout Twist1 gene and application of the disease model construction method in accurate treatment of inflammatory bowel disease, and the disease model construction method comprises the following steps: introducing a loxP site into the Twist1 gene through a CRISPR-Cas9 gene editing technology to generate a Twist1f / f mouse which can be conditionally knocked out; the method comprises the following steps: firstly, designing gRNA aiming at Twist1 and a donor vector containing loxP sites, and injecting the gRNA and the donor vector and Cas9 mRNA into a mouse fertilized egg together; mating the generated F0 generation with a wild type mouse to obtain an F1 generation Twist1f / f mouse; the F1 generation is mated with a transgenic mouse for expressing intestinal epithelium specific Cre recombinase, and finally a transgenic mouse (Twist1- / -) of which the intestinal epithelium specific knockout Twist1 is obtained; the knockout efficiency is verified through PCR, sequencing and protein expression analysis.

The application discloses application of PDGF-C in corneal injury repair. The application research finds that in a corneal injury model mouse in which Pdgf-c is knocked out, Pdgf-c knockout reduces the area of corneal healing in the corneal injury mouse model, and the number of corneal limbal stem cells with proliferative activity is significantly less than that of wild type mice. Through AAV-mediated PDGF-C overexpression, the proliferation of corneal limbal stem cells can be significantly promoted, and the area of corneal healing in the corneal injury mouse model is significantly increased. These results show that overexpression of PDGF-C significantly improves the ability of corneal injury healing. Therefore, the application provides a new method for treating corneal injury.

The invention relates to the technical field of biotechnology, in particular to a two-leaf aortic valve disease model mouse as well as a construction method and application thereof, and the construction method comprises the following steps: designing gRNA of a targeted mouse FGFR3 exon 5-7, adapting a T7 promoter, and carrying out transcription purification after off-target prediction; mixing gRNA with Cas9, adding glycerol, incubating to obtain an RNP compound, and verifying the activity; rNP is injected into fertilized eggs, and after culture, the fertilized eggs are transplanted to pseudopregnant female rats to obtain an F0 generation; the F0 and a wild type mouse are subjected to cage combination to obtain F1, F1 heterozygotes are subjected to reproduction to obtain F2, and homozygotes are identified through PCR and sequencing; and taking F2 heart HE staining ornamental deformity, and detecting the expression of FGFR3 by RT-PCR and WB. According to the model, R-N type two-leaf aortic valve and right ventricle double-outlet deformity can be stably reproduced, the construction method is accurate and efficient, the model can be used for disease mechanism research and drug screening, and the blank of specific subtype research tools is filled.

The invention relates to an mRNA (messenger ribonucleic acid) medicine for treating propionemia and application of the mRNA medicine. According to the invention, an optimal candidate mRNA molecule-PUMCH-MR-01 is selected by carrying out mRNA sequence optimization design on a propionyl-coenzyme A carboxylase alpha or beta (PCCA or PCCB) subunit amino acid sequence and carrying out in-vitro expression screening verification. Then, the protein expression level of PUMCH-MR-01 is evaluated in a wild type mouse body, and pharmacodynamic verification is carried out on cynomolgus monkeys. The result shows that the PUMCH-MR-01 can efficiently express the PCC enzyme protein, shows a certain drug effect trend in non-human primates, and can be used for effectively treating propionemia.

The invention belongs to the technical field of biology, and discloses a construction method and application of a Pik3ip1 gene knockout mouse model. The construction method comprises the following steps: (1) transferring sgRNA1 (SEQ ID No.2), sgRNA2 (SEQ ID No.3) and Cas9 protein into a mouse fertilized egg, and carrying out development and delivery to obtain an F0-generation chimeric mouse; (2) hybridizing the F0-generation chimeric mouse with a wild type mouse to obtain an F1-generation heterozygote mouse; and (3) performing same-genotype male and female mating on the F1-generation heterozygote mice to obtain the Pik3ip1 gene knockout mouse model. According to the invention, the Pik3ip1 gene knockout mouse model is constructed by utilizing the combination of sgRNA and CRISPR / Cas9 technology, the construction method is stable, and the constructed Pik3ip1 gene knockout mouse model can be applied to animal models such as inflammation and is used for deeply researching the action and mechanism of the Pik3ip1 gene in diseases such as inflammation.