CN101574074A - Model for dcf-1 gene knockout mouse, construction method and applications thereof - Google Patents
Model for dcf-1 gene knockout mouse, construction method and applications thereof Download PDFInfo
- Publication number
- CN101574074A CN101574074A CNA2009100533594A CN200910053359A CN101574074A CN 101574074 A CN101574074 A CN 101574074A CN A2009100533594 A CNA2009100533594 A CN A2009100533594A CN 200910053359 A CN200910053359 A CN 200910053359A CN 101574074 A CN101574074 A CN 101574074A
- Authority
- CN
- China
- Prior art keywords
- dcf
- gene knockout
- knockout mice
- gene
- mice model
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a gene dcf-1 related to the differentiation of neural stem cells and applications thereof. People obtain a dcf-1 gene knockout mouse by a homologous recombination, an embryonic stem cell transplantation and a Cre-loxP system, and find that the gene knockout can cause the mouse to generate anxiety; and by means of methods of realtime PCR, histochemistry and the like, people also find that the dcf-1 gene knockout can cause Pet-1 expression amount to be decreased, thus influencing the development of 5-serotonin neurons.
Description
Technical field
This patent relates to a kind of dcf-1 gene knockout mice model, its construction method and application thereof.
Background of invention
(Neural stem cell is that a class can be duplicated by self NSC) to neural stem cell, and has the cell that differentiation becomes neuron and spongiocyte potential.The high expressed of the dcf-1 gene relevant with Neural Stem Cells Differentiation helps stem cell to keep undifferentiated state, and the atomization that reticent dcf-1 expression of gene can the accelerans stem cell.
Mouse has become a kind of important model biology of research human diseases.The Cre-loxP system, thus knocking out of realization gene that can be more or less freely makes up the gene knockout mice model.Animal Behavior Science and histochemical developing rapidly, the new and high technology means of realtimePCR and so on are used in the laboratory, have given us the good platform from the angle research gene function of a biological integral.
Summary of the invention
One of purpose of the present invention is to provide a kind of dcf-1 gene knockout mice model relevant with Neural Stem Cells Differentiation.
Two of purpose of the present invention is to provide the construction method of this model.
Three of purpose of the present invention is to provide the application of this model in the screening anxiolytic drugs.
Four of purpose of the present invention is to provide the application of this model in preparation promotion serotonin neuronal development medicine.
For achieving the above object, the present invention adopts following technical scheme:
A kind of dcf-1 gene knockout mice model relevant with Neural Stem Cells Differentiation.
A kind of method that makes up above-mentioned dcf-1 gene knockout mice model is characterized in that the concrete steps of this method are: utilize the Cre-loxP system, targeting vector electric shock is transformed embryonic stem cell, make itself and genome generation homologous recombination.Again embryo stem cell transplantation is gone into the embryo, the both sides that are created in the dcf-1 gene have the offspring in loxP site, with these offsprings and the mouse hybridization of expressing the Cre recombinase, will produce dcf-1 gene knockout mice model.
The application of above-mentioned dcf-1 gene knockout mice model in the screening anxiolytic drugs.
The application of above-mentioned dcf-1 gene knockout mice model in preparation promotion serotonin neuronal development medicine.
The present invention is by the Cre-loxP system, and having made up really is reliable dcf-1 gene knockout mice model.By the Animal Behavior Science experiment, find that the rejecting of dcf-1 gene can cause mouse to produce anxiety then.For the better relation between research dcf-1 gene and the anxiety, the present invention has used means such as realtime round pcr and histochemistry, and the rejecting of further finding the dcf-1 gene can influence that serotonin is neuronic reaches maturity.
Description of drawings
Fig. 1 is a dcf-1 gene knockout mice design of graphics, indicates genome structure, authentication method and the qualification result of mouse among the figure.Wherein A-D is respectively the genome structure that wild-type mice, targeting vector, dcf-1 loxP mouse and dcf-1 reject mouse.E is the generation that confirms rejecting in genomic level.F is the generation that confirms rejecting at expression.
Fig. 2 is the behaviouristics experimental result of dcf-1 gene knockout mice, has shown the severe anxiety tendency.A is the number of times that enters open arms and middle section.B is the time that stops in open arms.
Fig. 3 is that dcf-1 gene knockout mice serotonin neuron is investigated the result, and the result shows that the serotonin neuronal quantity obviously reduces.A is the expression of serotonin neuron mark Pet-1, C-D be respectively wild type, dcf-1+/-and dcf-1-/-the nucleus raphes dorsalis section of mouse.
Embodiment
Embodiment one: the structure of dcf-1 gene knockout mice model
The rejecting of dcf-1 gene can utilize the Cre-loxP system to realize.Targeting vector electric shock is transformed embryonic stem cell, make itself and genome generation homologous recombination.Again embryo stem cell transplantation is gone into the embryo, the both sides that are created in the dcf-1 gene have the offspring in loxP site.With these offsprings and the mouse hybridization of expressing the Cre recombinase, will produce the dcf-1 gene knockout mice.By genome PCR, RT-PCR, the present invention has verified the disappearance that the dcf-1 gene has taken place in this model really, the result is as shown in Figure 1.
Embodiment two: the functional study of dcf-1 gene
1. behaviouristics research: the present invention carries out the behaviouristics test of overhead cross labyrinth to the dcf-1 gene knockout mice.Earlier mouse is placed on the middle section in labyrinth, observes the behavior of mouse in 5 minutes then, comprise number of times that enters open arms and middle section and the time that stops in the open arms zone.The result as shown in Figure 2, the WT mouse enter the number of times of middle section and dcf-1+/-and dcf-1-/-the not significantly difference of mouse, illustrate that the frequency that they shuttle back and forth is suitable substantially in the labyrinth, the chance of selecting to enter open arms and still be closure arm is also equal substantially; But wild type enters the number of times of open arms is higher than back two kinds of genotype, the WT mouse the time that open arms stops also obviously be longer than dcf-1+/-and dcf-1-/-mouse.The result shows: the behavior of dcf-1 gene knockout mice thing meet generally acknowledged anxiety behavioural characteristic unusually.
2.5-the hydroxytryptamine neuron is investigated: the present invention is directed to the neuronic mark Pet-1 of serotonin and carry out realtime PCR, brain tissue slice with the dcf-1 gene knockout mice also dyes simultaneously, the serotonin neuronal quantity of finding the dcf-1 gene knockout mice obviously reduces, and the result as shown in Figure 3.The result that neuron is investigated shows: the dcf-1 gene knockout causes mouse 5-HT neuron obviously to reduce, and is the reason that mouse produces the anxiety behavior.
Claims (4)
1. dcf-1 gene knockout mice model relevant with Neural Stem Cells Differentiation.
2. method that makes up dcf-1 gene knockout mice model according to claim 1 is characterized in that the concrete steps of this method are: utilize the Cre-loxP system, the targeting vector electric shock is transformed embryonic stem cell, make itself and genome generation homologous recombination.Again embryo stem cell transplantation is gone into the embryo, the both sides that are created in the dcf-1 gene have the offspring in loxP site, with these offsprings and the mouse hybridization of expressing the Cre recombinase, will produce dcf-1 gene knockout mice model.
3. the application of dcf-1 gene knockout mice model according to claim 1 in the screening anxiolytic drugs.
4. the application of dcf-1 gene knockout mice model according to claim 1 in preparation promotion serotonin neuronal development medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009100533594A CN101574074A (en) | 2009-06-18 | 2009-06-18 | Model for dcf-1 gene knockout mouse, construction method and applications thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009100533594A CN101574074A (en) | 2009-06-18 | 2009-06-18 | Model for dcf-1 gene knockout mouse, construction method and applications thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101574074A true CN101574074A (en) | 2009-11-11 |
Family
ID=41269254
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2009100533594A Pending CN101574074A (en) | 2009-06-18 | 2009-06-18 | Model for dcf-1 gene knockout mouse, construction method and applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101574074A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492686A (en) * | 2011-12-12 | 2012-06-13 | 湖南师范大学 | Kctd10 gene knock-out mouse model, construction method and application thereof |
| CN102766635A (en) * | 2012-05-25 | 2012-11-07 | 上海大学 | Construction method of human dcf1 gene transgenic drosophila melanogaster model |
| CN102925483A (en) * | 2012-05-25 | 2013-02-13 | 上海大学 | New uses of DCF1 gene |
| CN102940890A (en) * | 2012-10-11 | 2013-02-27 | 上海大学 | Gene application in inhibition and apoptosis of glioma cell |
| CN105974126A (en) * | 2016-05-31 | 2016-09-28 | 上海大学 | Application of dcf1 gene to regulation and control over expression of ATP1B1 |
-
2009
- 2009-06-18 CN CNA2009100533594A patent/CN101574074A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492686A (en) * | 2011-12-12 | 2012-06-13 | 湖南师范大学 | Kctd10 gene knock-out mouse model, construction method and application thereof |
| CN102766635A (en) * | 2012-05-25 | 2012-11-07 | 上海大学 | Construction method of human dcf1 gene transgenic drosophila melanogaster model |
| CN102925483A (en) * | 2012-05-25 | 2013-02-13 | 上海大学 | New uses of DCF1 gene |
| CN102766635B (en) * | 2012-05-25 | 2013-10-16 | 上海大学 | Construction method of human dcf1 gene transgenic drosophila melanogaster model |
| CN102940890A (en) * | 2012-10-11 | 2013-02-27 | 上海大学 | Gene application in inhibition and apoptosis of glioma cell |
| CN105974126A (en) * | 2016-05-31 | 2016-09-28 | 上海大学 | Application of dcf1 gene to regulation and control over expression of ATP1B1 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Schuman et al. | Four unique interneuron populations reside in neocortical layer 1 | |
| Ronce et al. | Dispersal syndromes | |
| Jiang et al. | Independent stem cell lineages regulate adipose organogenesis and adipose homeostasis | |
| Abbott et al. | Hybridization and speciation | |
| Qiu et al. | Circuit-specific intracortical hyperconnectivity in mice with deletion of the autism-associated Met receptor tyrosine kinase | |
| Putkey et al. | Unstable kinetochore-microtubule capture and chromosomal instability following deletion of CENP-E | |
| Low-Zeddies et al. | Chimera analysis of the Clock mutation in mice shows that complex cellular integration determines circadian behavior | |
| Anderson et al. | Reduced crossover interference and increased ZMM-independent recombination in the absence of Tel1/ATM | |
| CN101574074A (en) | Model for dcf-1 gene knockout mouse, construction method and applications thereof | |
| CN103179851B (en) | There is the Mus inflammatory model of IL33N terminal domains disappearance | |
| CN110951781B (en) | Construction method and application of epilepsia animal model | |
| Day et al. | Varied solutions to multicellularity: The biophysical and evolutionary consequences of diverse intercellular bonds | |
| JP5393492B2 (en) | Animal model, production method thereof, and drug candidate screening method using the same | |
| Kawakami et al. | Hybrid zones | |
| CN102492686B (en) | Kctd10 gene knock-out mouse model, construction method and application thereof | |
| CN108300737A (en) | A kind of method for building up and application thereof for the malignant lymphoma model that phenotype is highly consistent | |
| CN105440111B (en) | Pair of transcription activator-like effector nucleases (CTFs), coding sequences and application thereof | |
| Ramesh et al. | Can CRISPR help control locust populations to reduce the impact of plague outbreaks? | |
| US20180360005A1 (en) | Trpv1 knock-in method and trpv1 mice for adipose tissue modulation | |
| Abe et al. | A Cre knock‐in mouse line on the Sickle tail locus induces recombination in the notochord and intervertebral disks | |
| Behe | Theoretical Molecular Biology: Introductory Comments | |
| Bonthuis et al. | Dopa decarboxylase is a genetic hub of parental control over offspring behavior | |
| Kasimatis et al. | Sexual dimorphism through the lens of genome manipulation, forward genetics, and | |
| Chan | The Plasticity and Variation in Gene Expression during Development in C. elegans | |
| Carroll et al. | Functional genomics requires ecology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20091111 |