Stafford David A, Dichmann Darwin S, Chang Jessica K, Harland Richard M
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
Department of Genetics, Stanford University, Stanford, CA 94305-5120.
Proc Natl Acad Sci U S A. 2017 Jan 3;114(1):101-106. doi: 10.1073/pnas.1612069113. Epub 2016 Dec 16.
To define a complete catalog of the genes that are activated during mouse sclerotome formation, we sequenced RNA from embryonic mouse tissue directed to form sclerotome in culture. In addition to well-known early markers of sclerotome, such as Pax1, Pax9, and the Bapx2/Nkx3-2 homolog Nkx3-1, the long-noncoding RNA PEAT (Pax1 enhancer antisense transcript) was induced in sclerotome-directed samples. Strikingly, PEAT is located just upstream of the Pax1 gene. Using CRISPR/Cas9, we generated a mouse line bearing a complete deletion of the PEAT-transcribed unit. RNA-seq on PEAT mutant embryos showed that loss of PEAT modestly increases bone morphogenetic protein target gene expression and also elevates the expression of a large subset of ribosomal protein mRNAs.
为了确定在小鼠椎骨形成过程中被激活的基因的完整目录，我们对来自在培养中定向形成椎骨的胚胎小鼠组织的RNA进行了测序。除了众所周知的椎骨早期标志物，如Pax1、Pax9和Bapx2/Nkx3-2同源物Nkx3-1外，长链非编码RNA PEAT（Pax1增强子反义转录物）在定向形成椎骨的样本中被诱导。引人注目的是，PEAT位于Pax1基因的上游。利用CRISPR/Cas9，我们构建了一个完全缺失PEAT转录单元的小鼠品系。对PEAT突变胚胎进行的RNA测序显示，PEAT的缺失适度增加了骨形态发生蛋白靶基因的表达，同时也提高了很大一部分核糖体蛋白mRNA的表达。
