Department of Genetics, Harvard Medical School, Boston, MA, USA.
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA.
Science. 2018 Aug 31;361(6405). doi: 10.1126/science.aat9804. Epub 2018 Aug 9.
In vivo barcoding using nuclease-induced mutations is a powerful approach for recording biological information, including developmental lineages; however, its application in mammalian systems has been limited. We present in vivo barcoding in the mouse with multiple homing guide RNAs that each generate hundreds of mutant alleles and combine to produce an exponential diversity of barcodes. Activation upon conception and continued mutagenesis through gestation resulted in developmentally barcoded mice wherein information is recorded in lineage-specific mutations. We used these recordings for reliable post hoc reconstruction of the earliest lineages and investigation of axis development in the brain. Our results provide an enabling and versatile platform for in vivo barcoding and lineage tracing in a mammalian model system.
体内同源重组酶靶向突变标记是一种强大的记录生物信息的方法，包括发育谱系信息；然而，该方法在哺乳动物系统中的应用受到限制。我们利用多个归巢向导 RNA 在体内对小鼠进行标记，每个向导 RNA 可以产生数百个突变等位基因，组合后产生指数级多样性的条形码。这些向导 RNA 在受孕时被激活，并在妊娠期间持续产生突变，从而使胚胎发育过程中产生谱系特异性突变的标记。我们利用这些记录可靠地重建了最早的谱系，并研究了大脑中的轴突发育。我们的研究结果为哺乳动物模型系统中的体内条形码标记和谱系追踪提供了一个通用的有效平台。
