A specific synthetic RNA promotes cardiac myofibrillogenesis in the Mexican axolotl.
A specific synthetic RNA promotes cardiac myofibrillogenesis in the Mexican axolotl.
复制标题
一种特定的合成 RNA 促进墨西哥蝾螈的心肌原纤维形成。
DOI：
10.1006/bbrc.1996.1910
复制
发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Dube,DK
中科院分区：
文献类型：
--
作者：
Lemanski,LF;Nakatsugawa,M;Bhatia,R;Erginel-Unaltuna,N;Spinner,BJ;Dube,DK
墨西哥钝口螈是研究心脏发育的一种有趣的动物模型，因为它携带基因突变。纯合隐性 (c /c ) 突变胚胎的心脏不包含有组织的肌原纤维并且无法跳动。然而，这种缺陷可以通过在前内胚层 RNA 或内胚层中胚层条件培养基 RNA 存在下对突变心脏进行器官培养来纠正。我们从 pcDNAII 表达载体中的总条件培养基 RNA 构建了 cDNA 文库。我们通过器官培养生物测定筛选了 cDNA 文库，并分离出一个克隆 (Cl#4)，从中合成的 RNA 通过促进肌原纤维生成来纠正心脏缺陷。活性克隆的插入片段长度为 166 nt，具有独特的核苷酸序列。使用 SP6 RNA 聚合酶的 Cl#4 反义 RNA 未能拯救突变心脏。这种小 RNA 纠正突变心脏缺陷的能力表明该 RNA 可能不具有 mRNA 的作用。虽然确切的作用机制尚不清楚，但根据我们迄今为止的研究，非常清楚的是，Cl#4 RNA 的有义链具有促进肌原纤维生成并在体外和体内拯救突变心脏的能力。
Ambystoma mexicanumis an intriguing animal model for studying heart development because it carries a mutation in genec . Hearts of homozygous recessive (c /c ) mutant embryos do not contain organized myofibrils and fail to beat. However, the defect can be corrected by organ-culturing the mutant heart in the presence of RNA from anterior endoderm or RNA from endoderm mesoderm-conditioned medium. We constructed a cDNA library from total conditioned medium RNA in a pcDNAII expression vector. We screened the cDNA library by an organ culture bioassay and isolated a single clone (Cl#4), the synthetic RNA from which corrects the heart defect by promoting myofibrillogenesis. The insert size of the active clone is 166 nt in length with a unique nucleotide sequence. The anti-sense RNA from Cl#4 using SP6 RNA polymerase failed to rescue mutant hearts. The ability of this small RNA to correct the mutant heart defect suggests that the RNA probably does not act as an mRNA. While the precise mechanism of action is not yet known, on the basis of our studies to date it is very clear that the sense strand of Cl#4 RNA has the ability to promote myofibrillogenesis and rescue the mutant hearts bothin vitroandin vivo.