Application of an F0-based genetic assay in adult zebrafish to identify modifier genes of an inherited cardiomyopathy.
Application of an F0-based genetic assay in adult zebrafish to identify modifier genes of an inherited cardiomyopathy.
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DOI：
10.1242/dmm.049427
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发表时间：
2023-05-01
影响因子：
4.3
通讯作者：
Xu, Xiaolei
中科院分区：
文献类型：
--
作者：
Ding, Yonghe;Wang, Mingmin;Bu, Haisong;Li, Jiarong;Lin, Xueying;Xu, Xiaolei
修饰基因对我们理解人类疾病的病理生理学有很大的贡献；然而，识别修饰基因的有效方法仍然缺乏。在这里，我们的目标是以bag3基因敲除(bag3e2/e2)心肌病模型为范例，在成年斑马鱼中建立基于F0的快速遗传检测方法。首先，通过利用经典的遗传育种方法，我们鉴定了dnajb6b是bag3心肌病的有害修饰基因。接下来，我们通过注射预测的微同源介导的末端连接(MMEJ)诱导的单引导RNA/Cas9蛋白复合体，在成年斑马鱼中建立了基于F0的遗传检测方法。我们发现在F0成鱼中保持了有效的基因敲除，使mTOR单倍体不足的有益修饰效应和dna jb6b基因对bag3心肌病的有害修饰效应得以重现。最后，我们采用了基于F0的遗传分析方法来筛选在bag3心肌病模型中差异表达的基因。因此，myh9b被鉴定为一个新的bag3心肌病修饰基因。总而言之，这些数据证明了基于F0成年斑马鱼的遗传检测的可行性，该检测可以有效地用于发现遗传性心肌病的修饰基因。摘要：由于修饰基因的原因，许多含有相同原因突变的人类疾病的表型可能有很大的差异。使用BAG3相关的心肌病斑马鱼模型，我们证明了一种基于F0的遗传分析的可行性，该方法能够快速发现疾病修饰物。
Modifier genes contribute significantly to our understanding of pathophysiology in human diseases; however, effective approaches to identify modifier genes are still lacking. Here, we aim to develop a rapid F0-based genetic assay in adult zebrafish using the bag3 gene knockout (bag3e2/e2) cardiomyopathy model as a paradigm. First, by utilizing a classic genetic breeding approach, we identified dnajb6b as a deleterious modifier gene for bag3 cardiomyopathy. Next, we established an F0-based genetic assay in adult zebrafish through injection of predicted microhomology-mediated end joining (MMEJ)-inducing single guide RNA/Cas9 protein complex. We showed that effective gene knockdown is maintained in F0 adult fish, enabling recapitulation of both salutary modifying effects of the mtor haploinsufficiency and deleterious modifying effects of the dnajb6b gene on bag3 cardiomyopathy. We finally deployed the F0-based genetic assay to screen differentially expressed genes in the bag3 cardiomyopathy model. As a result, myh9b was identified as a novel modifier gene for bag3 cardiomyopathy. Together, these data prove the feasibility of an F0 adult zebrafish-based genetic assay that can be effectively used to discover modifier genes for inherited cardiomyopathy. Summary: Owing to modifier genes, phenotypes of many human diseases that harbor identical causal mutations can vary substantially. Using a BAG3-associated cardiomyopathy zebrafish model, we demonstrated the feasibility of a F0-based genetic assay that enables rapid discovery of disease modifiers.
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影响因子：
20.1
作者：
Ding Y;Sun X;Huang W;Hoage T;Redfield M;Kushwaha S;Sivasubbu S;Lin X;Ekker S;Xu X
通讯作者：
Xu X
影响因子：
5.4
作者：
Ding Y;Yang J;Chen P;Lu T;Jiao K;Tester DJ;Giudicessi JR;Jiang K;Ackerman MJ;Li Y;Wang DW;Lee HC;Wang DW;Xu X
通讯作者：
Xu X
DOI：
10.1016/j.jacbts.2017.09.009
发表时间：
2018-03
期刊：
JACC. Basic to translational science
影响因子：
--
作者：
Myers VD;McClung JM;Wang J;Tahrir FG;Gupta MK;Gordon J;Kontos CH;Khalili K;Cheung JY;Feldman AM
通讯作者：
Feldman AM
影响因子：
9.8
作者：
Norton, Nadine;Li, Duanxiang;Hershberger, Ray E.
通讯作者：
Hershberger, Ray E.
影响因子：
37.8
作者：
Goldberg IJ;Reue K;Abumrad NA;Bickel PE;Cohen S;Fisher EA;Galis ZS;Granneman JG;Lewandowski ED;Murphy R;Olive M;Schaffer JE;Schwartz-Longacre L;Shulman GI;Walther TC;Chen J
通讯作者：
Chen J