疯狂做受xxxx高潮不断,亚洲中文无码永久免弗,亚洲免费视频免在线观看,国产老妇伦国产熟女老妇高清

基因編輯斑馬魚(yú)

南模生物斑馬魚(yú)技術(shù)服務(wù)平臺(tái)擁有受過(guò)嚴(yán)格訓(xùn)練的技術(shù)與分析人員，可提供高質(zhì)量的斑馬魚(yú)基因編輯服務(wù)，并可通過(guò)分析基因敲除、敲低、過(guò)表達(dá)斑馬魚(yú)模型的表型改變研究基因、蛋白在體內(nèi)的功能以及相關(guān)信號(hào)通路，篩選致病基因、探索基因功能。

我們提供以下服務(wù)：

基因Knock-down及表型分析（Morpholino knock-down）
基因Over-expression及表型分析
基因敲除斑馬魚(yú)定制（Cas9-KO）
轉(zhuǎn)基因斑馬魚(yú)定制

聯(lián)系我們了解更多服務(wù)詳情，歡迎來(lái)電：400-728-0660，或來(lái)函 info@modelorg.com。

基因Knock-down及表型分析（Morpholino knock-down）

Morpholino knock-down技術(shù)原理

圖1.?Morpholino 技術(shù)原理。（a）Morpholino與RNA結(jié)合。（b）Morpholino 通過(guò)阻斷翻譯過(guò)程而發(fā)揮作用。（c-e）Morpholino阻斷RNA的正常剪接。

Morpholino Knockdown應(yīng)用案例

圖2.?Morpholino Knockdown 靶點(diǎn)位置示意圖。The zebrafish gene-X was targeted by three specific morpholino antisense strategies to prevent either the translation of the zebrafish gene (ATG-MO) or proper splicing of exon 3 (E3I3-MO).?

Morpholino Knockdown表型分析

圖3.?Panels A through H show lateral views of control MO injected zebrafish embryos (Panel A and Panel E) and embryos injected with? gene-X morpholino oligonucleotides (MO) (Panel B through C, Panel F through G), gene-X-e3i3-MO plus nonmutant zebrafish gene-X? (Panel D and Panel H). Coinjection of nonmutant zebrafish gene-X mRNA rescued U-shaped somites (red arrow) and curved body axis? (blue dotted line) in gene-X morphants at 52 hpf. The bar graph in Panel I shows the percentage of embryos with development defects.Panels J Effectiveness of gene-X knockdown was confirmed by RT-PCR and sanger sequencing. hpf, hours post fertilization.?

Morpholino Knockdown信號(hào)通路機(jī)制分析

圖4.? Endogenous shha, ptch1, ptch2, sufu, gli1, gli2a, gli2b, and gli3 in wild-type control and geneX morphants assessed by qRT-PCR (n = 100 individual embryos). geneX fine-tunes Hedgehog patterning activity maybe through a novel regulatory feedback loop.?

基因Knockdown后抑制斑馬魚(yú)血管生成

圖5.?GeneX? knock down inhibits the trunk angiogenesis in zebrafish. (A-D) Representative fluorescent images of zebrafish embryos at 32h post-fertilization (hpf). (C-E) Compared with wild-type control, embryos injected with geneX-MO present a lower number of incomplete ISVs and only occasional sprouts (asterisk) of dorsal aorta. The boxed regions are shown at higher magnification in the right panels. DLAV, dorsal longitudinal anastomotic vessels; ISV, intersegmental vessels; DA, dorsal aorta; PCV, posterior cardinal vein.??

基因Knockdown后導(dǎo)致中樞神經(jīng)系統(tǒng)特異性細(xì)胞凋亡

圖6. Morpholino knock down of geneX induces potent CNS-specific apoptosis. Wild-type control embryos and embryos injected with geneX-MO were stained with acridine orange (AO) at 32hpf. Apoptotic cells are visible as black spots, and less bright homogenous black staining is unspecific background staining. (A-B) Uninjected wild-type control zebrafish exhibited few or no apoptotic cells in CNS (central nervous system). In contrast, significantly increased staining was observed throughout? the CNS in zebrafish injected with geneX-MO (C-D). The blue boxed regions are shown at higher magnification in the right panels. A-D: lateral view, anterior, left.?

基因Knockdown后具有潛在聽(tīng)毒性

圖7. GeneX knock down induces potent ototoxicity in zebrafish. Wild-type control embryos and embryos injected with geneX-MO were stained with the mitochondrial potentiometric dye DASPEI at 6-dpf. Hair cells stereotypically located on the lateral line were stained as green dots (white arrow). Uninjected wild-type control zebrafish exhibited normal hair cell number. In contrast, significantly decreased hair cell staining was observed in zebrafish injected with geneX-MO. Fluorescent DASPEI images were inverted for particle analysis. The fluorescence particle signal was quantified using morphometric analysis. dpf, days post fertilization.?

文獻(xiàn)發(fā)表

Wang MS, Zhang RW, Su LY, et al. Positive selection rather than relaxation of functional constraint drives the evolution of vision duringchicken domestication. Cell Res. 2016 May;26(5):556-73.?

基因Over-expression及表型分析

482f54775de80eb785392a44f5d49846副本.png

Over-expression 應(yīng)用案例

圖8. Nonmutant bmp10 overexpression causes development defects in zebrafish.? (A-C) Gross morphology at 32hpf. Compared with uninjected wild-type control embryos, nonmutant zebrafish bmp10 overexpression causes decreased body length (black dotted line) and curved body axis (blue dotted line) in zebrafish . The bar graph in Panel D through E show the percentage and body length of embryos with development defects. hpf, hours post fertilization.?

圖9. Nonmutant bmp10 overexpression inhibits the trunk angiogenesis in zebrafish.? (A-F) Representative bright field and fluorescent images of zebrafish embryos at 32h post-fertilization (hpf). Red arrow indicates haemorrhage in the tail (B). (C-H) Compared with wild-type control, nonmutant? zebrafish bmp10 mRNA (200pg) injection present a lower number of incomplete ISVs and only occasional sprouts (asterisk) of dorsal aorta. The red boxed regions are shown at higher magnification in the right panels. DLAV, dorsal longitudinal anastomotic vessels; ISV, intersegmental vessels; DA, dorsal aorta; PCV, posterior cardinal vein.?

文獻(xiàn)發(fā)表

Wang MS, Huo YX, Li Y, et al. Comparative population genomics reveals genetic basis underlying body size of domestic chickens.J Mol Cell Biol. 2016 Dec;8(6):542-552.?
Feng N, Chen H, Fu S, et al. HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed bythalidomide. Sci Rep. 2016 Jun 1;6:27280.??

基因敲除斑馬魚(yú)定制（Cas9-KO）

利用CRISPR/Cas9技術(shù)，針對(duì)靶基因序列設(shè)計(jì)sgRNA，指導(dǎo)Cas9蛋白在特定基因位點(diǎn)引起DNA雙鏈斷裂，在非同源性末端接合修復(fù)斷裂DNA的過(guò)程中，靶基因堿基突變或缺失被引入到斑馬魚(yú)基因組中，最終導(dǎo)致靶基因無(wú)法正常轉(zhuǎn)錄翻譯，達(dá)到基因敲除的目的。目前我們利用CRISPR-Cas9技術(shù)，提供AB品系的基因敲除斑馬魚(yú)制備，AB系是最為普遍使用的標(biāo)準(zhǔn)純遺傳背景品系之一。同時(shí)也可根據(jù)委托方的需求，提供TU、Casper 以及其它遺傳背景的基因敲除斑馬魚(yú)服務(wù)。