Application of the 3C Method to Study the Developmental Genes in Drosophila Larvae

被引:1
作者
Bylino, Oleg V. [1 ]
Ibragimov, Airat N. [1 ,2 ]
Digilio, Filomena Anna [3 ]
Giordano, Ennio [4 ]
Shidlovskii, Yulii V. [1 ,5 ]
机构
[1] Russian Acad Sci, Inst Gene Biol, Dept Gene Express Regulat Dev, Moscow, Russia
[2] Russian Acad Sci, Inst Gene Biol, Ctr Precis Genome Editing & Genet Technol Biomed, Moscow, Russia
[3] UOS Naples CNR, Res Inst Terr Ecosyst IRET, Naples, Italy
[4] Univ Napoli Federico II, Dept Biol, Naples, Italy
[5] IM Sechenov First Moscow State Med Univ, Dept Biol & Gen Genet, Moscow, Russia
基金
俄罗斯科学基金会; 俄罗斯基础研究基金会;
关键词
chromatin conformation capture; distal interaction; larvae; chromatin; enhancer; promoter; Drosophila; CHROMOSOME CONFORMATION CAPTURE; HIGH-RESOLUTION; CHROMATIN INTERACTIONS; HI-C; SINGLE-CELL; SACCHAROMYCES-CEREVISIAE; PROMOTER CONTACTS; HSP70; PROMOTER; SNF6; PROTEINS; ENHANCER;
D O I
10.3389/fgene.2022.734208
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
A transition from one developmental stage to another is accompanied by activation of developmental programs and corresponding gene ensembles. Changes in the spatial conformation of the corresponding loci are associated with this activation and can be investigated with the help of the Chromosome Conformation Capture (3C) methodology. Application of 3C to specific developmental stages is a sophisticated task. Here, we describe the use of the 3C method to study the spatial organization of developmental loci in Drosophila larvae. We critically analyzed the existing protocols and offered our own solutions and the optimized protocol to overcome limitations. To demonstrate the efficiency of our procedure, we studied the spatial organization of the developmental locus Dad in 3rd instar Drosophila larvae. Differences in locus conformation were found between embryonic cells and living wild-type larvae. We also observed the establishment of novel regulatory interactions in the presence of an adjacent transgene upon activation of its expression in larvae. Our work fills the gap in the application of the 3C method to Drosophila larvae and provides a useful guide for establishing 3C on an animal model.
引用
收藏
页数:19
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