研究人员使用Crispr可以敲除 一种叫作CCR5的基因。 By using CRISPR, He was able to delete a gene known as CCR5. 敲除 Rubicon会增强自噬和内吞转运(11,12)。Knockdown of Rubicon enhances autophagy and endocytic trafficking(11,12).案例一基因组规模的CRISPR-Cas9敲除 和转录激活筛选. Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. 小鼠ESC中Tcf3敲除 可引起分化延迟,上调Nanog[15]。 Tcf3 knockdown in mouse ESCs can cause delayed differentiation and upregulate Nanog[15]. KuiLiu和他的同事在特异组织敲除 的小鼠上开展研究实验。 Kui Liu and his colleagues performed experiments on tissue-specific knockout mice.
在之前,如果我们敲除 一个基因,我们仅能够在第一步观察这会产生何种影响。 Before, if we knocked out a gene, we could only see what effect this had at the very first step. 敲除 alr(mll2)可揭示alr靶基因并导致细胞粘附和生长的改变。Knockdown of ALR(MLL2) reveals ALR target genes and leads to alterations in cell adhesion and growth". 之前,如果我们敲除 一个基因,我们只能在第一步观察到这将带来怎样的影响。 Before, if we knocked out a gene, we could only see what effect this had at the very first step. 奇怪的控制小鼠漫游,而敲除 小鼠经常粘在一个小区域,并重复相同的路线。 Curious control mice roamed while knockout mice often stuck to a small area and repeated the same routes. 但是CRISPR可以敲除 或者精确地编辑DNA序列而不会引入任何外来的DNA。 But CRISPR can knockout or precisely edit DNA sequences without leaving behind any foreign DNA. 敲除 大脑FNDC5/irisin会损害小鼠的长时程增强和新的物体识别记忆。Knockdown of brain FNDC5/irisin impairs long-term potentiation and novel object recognition memory in mice. 或者,切割可以同时“敲除 ”不需要的基因,例如,导致疾病的基因。 Alternatively, the cut can altogether“knock out ” of a particular unwanted gene- for example, a gene that causes diseases. Chai说,敲除 一个调节因子可以致癌,但调节其对立的调节因子可以阻止癌症。 Knocking out one regulator can create cancer, Chai said, but modulating its opposing regulator can stop the cancer. 但在这项研究中,研究人员通过基因敲除 NMDA受体来了解其作用可能是什么。 But in this study, the researchers genetically knocked out the NMDA receptor to understand what their role might be. 为了测试这个想法,该团队尝试了几年制造缺乏miR-140的靶向“敲除 ”小鼠模型。 To test this idea, the team tried for several years to make targeted“knockout ” mouse models that lacked miR-140. 如果这些研究人员抑制IL-17a或敲除 IL-17a受体,那么这种症状逆转不会发生。 If the researchers inhibited IL-17a or knocked out the receptors for IL-17a, this symptom reversal did not occur. 令人惊讶的是,敲除 试验阐释了c-FLIP在CD95信号中能够抑制NF-κB激活。 Surprisingly, knockdown experiments revealed that c-FLIP inhibits NF-κB activation in the context of CD95 signaling. 敲除 一种叫做ATG5的蛋白质之后(它对自噬过程至关重要),细胞凋亡率也会提高。Knocking out a protein called ATG5, which is critical to the autophagy process, also increased the rate of apoptosis. 研究人员发现当他们敲除 小鼠前列腺癌模型中的HK2后,这些小鼠身上的肿瘤停止生长。 The researchers found that when they knocked out HK2 in a mouse prostate cancer model, the tumors in these mice stopped growing. 同时,完全敲除 cathepsinB基因的另一项实验也成功使线虫的生育力扩大了约10%。 Another experiment that knocked out the cathepsin B genes entirely succeeded in extending worms' fertility by about 10 percent. 在我们的最新作品,我们通过使用基因“敲除 ”细菌来测试这种细菌,这种细菌已经被设计成仅失去一个泵基因。 In our latest work, we tested this by using gene“knockout ” bacteria, which had been engineered to lose just one pump gene. 相反,Fabp7基因敲除 小鼠,表现出多动和焦虑,这是精神分裂症患者中观察到的类似表型。 In contrast, FABP7 knock-out mice displayed hyperactivity and anxiety, a phenotype similar to that observed in schizophrenic patients. 在过去的4年中,CRISPR/Cas9靶向基因敲除 已迅速成为植物研究中最流行的反向遗传学研究工具。 In the past years, CRISPR/ Cas9 targeted gene knockout has rapidly become the most popular reverse genetics research tool in plant research. 相比之下,通过敲除 基因的方式阻断PD-1确定性更强;同时,细胞增殖能够增加响应的机会。 By contrast, knocking out the gene blocks PD-1 with greater certainty, while multiplying the cells increases the chance of a response. 研究人员可使用Lipofectamine™2000试剂进行基于siRNA和shRNA的基因敲除 实验及基因表达研究。 Researchers use Lipofectamine® 2000 Reagent for siRNA- and shRNA-based gene knockdown experiments, as well as for gene expression studies. 研究者可利用RNAi快速“敲除 ”特定基因的表达,从而可以确定这些基因的生物学功能。 RNAi allows researchers to rapidly“knock out ” the expression of specific genes and, thus, to define the biological functions of those genes. 当我们将ythdf1敲除 与pd-l1抗体联合使用时,我们几乎控制住了所有小鼠的肿瘤,”何川说[3]。 Once we combined YTHDF1 knock-out with the checkpoint inhibitor anti-PD-L1, we got almost complete tumor control in a mouse model,” He explained. 通过直接筛选蛋白产生,可以确认基因敲除 ,避免与蛋白功能无关的非特异性阳性信号(假阳性)。 By screening directly for protein production, genetic knockout can be confirmed, and non-specific positive signals(false positives) unrelated to protein function can be avoided. 通过敲除 或突变线虫基因,观察其效应,研究人员能够了解一些基因影响动物结构或生理学的机制。 By knocking out or mutating roundworm genes and observing the effects, researchers can obtain insight into how genes influence the animal's structure or physiology. 当研究人员检查具有基质金属蛋白酶或pannexin1敲除 的小鼠时,他们发现Casp11突变可能导致这些动物对休克的抵抗。 When the researchers examined mice with matrix metalloproteinases or pannexin 1 knocked out , they found that Casp11 mutations likely contributed to these animals' resistance to shock.
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