College of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY 12203, USA.
Exp Biol Med (Maywood). 2022 Dec;247(23):2119-2127. doi: 10.1177/15353702221121600. Epub 2022 Sep 16.
DNA barcoding is a method to identify biological entities, including individual cells, tissues, organs, or species, by unique DNA sequences. With the advent of next generation sequencing (NGS), there has been an exponential increase in data acquisition pertaining to medical diagnosis, genetics, toxicology, ecology, cancer, and developmental biology. While barcoding first gained wide access in identifying species, signature tagged mutagenesis has been useful in elucidating gene function, particularly in microbes. With the advent of CRISPR/CAS9, methodology to profile eukaryotic genes has made a broad impact in toxicology and cancer biology. Designed homing guide RNAs (hgRNAs) that self-target DNA sequences facilitate cell lineage barcoding by introducing stochastic mutations within cell identifiers. While each of these applications has their limitations, the potential of sequence barcoding has yet to be realized. This review will focus on signature-tagged mutagenesis and briefly discuss the history of barcoding, experimental problems, novel detection methods, and future directions.
DNA 条形码是一种通过独特的 DNA 序列来识别生物实体的方法,包括单个细胞、组织、器官或物种。随着下一代测序(NGS)的出现,与医学诊断、遗传学、毒理学、生态学、癌症和发育生物学相关的数据获取呈指数级增长。虽然条形码最初广泛用于识别物种,但标记突变体技术在阐明基因功能方面非常有用,特别是在微生物中。随着 CRISPR/CAS9 的出现,用于描绘真核基因的方法在毒理学和癌症生物学中产生了广泛的影响。设计的同源定向 guide RNA(hgRNA)通过在细胞标识符内引入随机突变,促进真核细胞的谱系条形码。虽然这些应用都有其局限性,但序列条形码的潜力尚未实现。这篇综述将重点介绍标记突变体技术,并简要讨论条形码的历史、实验问题、新的检测方法和未来方向。