Deterministic assembly of chromosome ensembles in a programmable membrane trap array.

Chromosomes Microfluidic Analytical Techniques Microfluidics
chromosome manipulation droplet synthetic biology

Journal

Biofabrication
ISSN: 1758-5090
Titre abrégé: Biofabrication
Pays: England
ID NLM: 101521964

Informations de publication

Date de publication:
20 07 2021
Historique:
received: 17 04 2021
accepted: 07 07 2021
pubmed: 8 7 2021
medline: 10 11 2021
entrez: 7 7 2021
Statut: epublish

Résumé

Selective spatial isolation and manipulation of single chromosomes and the controlled formation of defined chromosome ensembles in a droplet-based microfluidic system is presented. The multifunctional microfluidic technology employs elastomer valves and membrane displacement traps to support deterministic manipulation of individual droplets. Picoliter droplets are formed in the 2D array of microscale traps by self-discretization of a nanoliter sample plug, with membranes positioned over each trap allowing controllable metering or full release of selected droplets. By combining discretization, optical interrogation, and selective droplet release for sequential delivery to a downstream merging zone, the system enables efficient manipulation of multiple chromosomes into a defined ensemble with single macromolecule resolution. Key design and operational parameters are explored, and co-compartmentalization of three chromosome pairs is demonstrated as a first step toward formation of precisely defined chromosome ensembles for applications in genetic engineering and synthetic biology.

Identifiants

DOI: 10.1088/1758-5090/ac1258 PMID: 34233304 PMC: PMC9974010
pubmed: 34233304
doi: 10.1088/1758-5090/ac1258
pmc: PMC9974010
mid: NIHMS1869219
doi:

Types de publication

Journal Article Research Support, N.I.H., Intramural Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : Intramural NIH HHS
ID : Z01 BC010309
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA BC010833
Pays : United States

Informations de copyright

© 2021 IOP Publishing Ltd.

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Auteurs

Hesam Babahosseini (H)

National Cancer Institute, National Institutes of Health, Bethesda, MD, United States of America.
Department of Mechanical Engineering, University of Maryland, College Park, MD, United States of America.

Darawalee Wangsa (D)

National Cancer Institute, National Institutes of Health, Bethesda, MD, United States of America.

Mani Pabba (M)

Department of Mechanical Engineering, University of Maryland, College Park, MD, United States of America.

Thomas Ried (T)

National Cancer Institute, National Institutes of Health, Bethesda, MD, United States of America.

Tom Misteli (T)

National Cancer Institute, National Institutes of Health, Bethesda, MD, United States of America.

Don L DeVoe (DL)

Department of Mechanical Engineering, University of Maryland, College Park, MD, United States of America.
ORCID: 0000-0002-7740-9993

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Classifications MeSH

questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Chromosomes › Deterministic assembly of chromosome ensembles...
questionsmedicales.fr › Techniques d'investigation › Procédures d'analyse sur micropuce › Techniques d'analyse microfluidique › Deterministic assembly of chromosome ensembles...
questionsmedicales.fr › Technologie, industrie et agriculture › Technologie › Miniaturisation › Microtechnologie › Microfluidique › Deterministic assembly of chromosome ensembles...