γ CaMKII 的遗传补偿，一个进化上保守的基因。

Genetic compensation of γ CaMKII, an evolutionarily conserved gene.

机构信息

Life Sciences, Virginia Commonwealth University, Richmond, VA 23284, USA.

Department of Cell Biology, University of Virginia, Charlottesville, VA, 22903, USA; Department of Biotechnology and Bioindustry Science, National Cheng-Kung University, Tainan 70101, Taiwan; The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.

出版信息

Gene. 2020 Jun 5;742:144567. doi: 10.1016/j.gene.2020.144567. Epub 2020 Mar 10.

DOI:10.1016/j.gene.2020.144567

PMID:32165308

Abstract

CaMKII is a Ca/CaM-dependent protein kinase encoded by a family of conserved genes found throughout all metazoan species and expressed from fertilization into adulthood. One of these genes, camk2g1, is particularly important during early development as determined by pharmacologic, dominant negative and antisense morpholino approaches in zebrafish. Four other teleost fish species (cavefish, medaka, stickleback, and tilapia), exhibit sequence conservation of camk2g1 and duplication of the same CaMKII genes. A homozygous mutant of camk2g1 was generated in zebrafish using TALEN technology but yielded none of the phenotypic alterations seen using all other approaches and was reproductively viable. However, these camk2g1 mutant embryos showed a 4-fold over-expression of its paralog camk2g2. None of the other camk2 genes showed such transcriptional elevation, in fact, some of these genes were suppressed to 10% of wild type levels. In contrast, G0 camk2g1 CRISPR/Cas9 embryos recapitulated nearly all of the altered phenotypes observed in camk2g1 morphants, including renal, aural and ciliary defects. These findings validate the importance of this gene family during early zebrafish development and provide evidence for gene-specific transcriptional cross-talk consistent with genetic compensation.

摘要

钙调蛋白依赖性蛋白激酶 II（CaMKII）是一种由在所有后生动物物种中发现的保守基因家族编码的 Ca/CaM 依赖性蛋白激酶，从受精到成年期都有表达。这些基因中的一个，camk2g1，在斑马鱼中通过药理学、显性负性和反义 morpholino 方法确定在早期发育中尤为重要。另外四种硬骨鱼物种（洞穴鱼、日本青鳉、棘鱼和罗非鱼），camk2g1 的序列保守性和相同的 CaMKII 基因的复制。使用 TALEN 技术在斑马鱼中产生了 camk2g1 的纯合突变体，但没有产生使用所有其他方法观察到的表型改变，并且具有生殖能力。然而，这些 camk2g1 突变体胚胎显示其旁系同源物 camk2g2 的表达增加了 4 倍。没有其他 camk2 基因表现出这种转录水平的升高，事实上，其中一些基因被抑制到野生型水平的 10%。相比之下，G0 camk2g1 CRISPR/Cas9 胚胎几乎重现了在 camk2g1 morphants 中观察到的所有改变的表型，包括肾脏、听觉和纤毛缺陷。这些发现验证了该基因家族在早期斑马鱼发育中的重要性，并提供了证据表明存在基因特异性转录交叉对话，与遗传补偿一致。