生物
基因
基因组
计算生物学
遗传学
人类基因组
DNA
功能基因组学
分子生物学
基因组学
作者
Leslie A. Mitchell,Laura H. McCulloch,Sudarshan Pinglay,Henri Berger,Nazario Bosco,Ran Brosh,Milica Bulajić,Emily Huang,Megan S. Hogan,James A. Martin,Esteban O. Mazzoni,Teresa Davoli,Matthew T. Maurano,Jef D. Boeke
出处
期刊:Genetics
[Oxford University Press]
日期:2021-03-20
卷期号:218 (1)
被引量:27
标识
DOI:10.1093/genetics/iyab038
摘要
Design and large-scale synthesis of DNA has been applied to the functional study of viral and microbial genomes. New and expanded technology development is required to unlock the transformative potential of such bottom-up approaches to the study of larger mammalian genomes. Two major challenges include assembling and delivering long DNA sequences. Here, we describe a workflow for de novo DNA assembly and delivery that enables functional evaluation of mammalian genes on the length scale of 100 kilobase pairs (kb). The DNA assembly step is supported by an integrated robotic workcell. We demonstrate assembly of the 101 kb human HPRT1 gene in yeast from 3 kb building blocks, precision delivery of the resulting construct to mouse embryonic stem cells, and subsequent expression of the human protein from its full-length human gene in mouse cells. This workflow provides a framework for mammalian genome writing. We envision utility in producing designer variants of human genes linked to disease and their delivery and functional analysis in cell culture or animal models.
科研通智能强力驱动
Strongly Powered by AbleSci AI