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Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors.

Title: Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors.
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Name(s): Sima, Jiao, author
Bartlett, Daniel A, author
Gordon, Molly R, author
Gilbert, David M, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2018-02-28
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: The role of DNA sequence in determining replication timing (RT) and chromatin higher order organization remains elusive. To address this question, we have developed an extra-chromosomal replication system (E-BACs) consisting of ∼200 kb human bacterial artificial chromosomes (BACs) modified with Epstein-Barr virus (EBV) stable segregation elements. E-BACs were stably maintained as autonomous mini-chromosomes in EBNA1-expressing HeLa or human induced pluripotent stem cells (hiPSCs) and established distinct RT patterns. An E-BAC harboring an early replicating chromosomal region replicated early during S phase, while E-BACs derived from RT transition regions (TTRs) and late replicating regions replicated in mid to late S phase. Analysis of E-BAC interactions with cellular chromatin (4C-seq) revealed that the early replicating E-BAC interacted broadly throughout the genome and preferentially with the early replicating compartment of the nucleus. In contrast, mid- to late-replicating E-BACs interacted with more specific late replicating chromosomal segments, some of which were shared between different E-BACs. Together, we describe a versatile system in which to study the structure and function of chromosomal segments that are stably maintained separately from the influence of cellular chromosome context.
Identifier: FSU_pmch_29294101 (IID), 10.1093/nar/gkx1265 (DOI), PMC5829748 (PMCID), 29294101 (RID), 29294101 (EID), 4774271 (PII)
Grant Number: P01 GM085354, R01 GM083337
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5829748.
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_29294101
Host Institution: FSU
Is Part Of: Nucleic acids research.
1362-4962
Issue: iss. 4, vol. 46

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Sima, J., Bartlett, D. A., Gordon, M. R., & Gilbert, D. M. (2018). Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors. Nucleic Acids Research. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_29294101