Abstract/Details

An inside-out approach to nuclear mechanics: Genetic engineering of an in vitro laminopathy model

Thompson, Matthew.   Purdue University ProQuest Dissertations & Theses,  2016. 10148463.

Abstract (summary)

Laminopathies are a group of genetic diseases affecting the nuclear lamina of metazoan cells with mutations in the genes LMNA, LMNB1, and LMNB2, which code for intermediate filament proteins called lamins. In LMNA alone, more than 180 mutations causing at least 14 diseases exist displaying a variety of phenotypes. Classical Hutchinson-Gilford Progeria Syndrome (HGPS) is a progeroid, or accelerated aging, laminopathy caused by a de novo heterozygous point mutation in LMNA (c. 1824 C>T). The disease affects multiple organ systems resulting in apparent accelerated aging and early death due to atherosclerotic complications such as myocardial infarction or stroke. At the subcellular level, HGPS causes stiffening of the nuclear envelope, an increase in mechanosensitivity, inhibited DNA repair, aberrant chromatin organization, and other complications. Here, the CRISPR-Cas9 genome editing system has been used to successfully incorporate the HGPS mutation into the human HT1080 fibroblast cell line using paired D10A mutant Cas9 nickases guided by truncated guide RNAs (tru-gRNAs) with low levels of off-target mutagenesis. This in vitro genetic disease model may be used in nuclear mechanics studies of HGPS to further uncover disease mechanisms and potential treatments or in basic science research aimed at learning the roles played by nuclear lamins and their mutant forms.

Indexing (details)


Subject
Genetics;
Biomedical engineering
Classification
0369: Genetics
0541: Biomedical engineering
Identifier / keyword
Biological sciences; Applied sciences
Title
An inside-out approach to nuclear mechanics: Genetic engineering of an in vitro laminopathy model
Author
Thompson, Matthew
Number of pages
61
Degree date
2016
School code
0183
Source
MAI 56/01M(E), Masters Abstracts International
ISBN
978-1-369-03644-2
Advisor
Neu, Corey P.
Committee member
Calve, Sarah; Umulis, David M.
University/institution
Purdue University
Department
Biomedical Engineering
University location
United States -- Indiana
Degree
M.S.B.M.E.
Source type
Dissertation or Thesis
Language
English
Document type
Dissertation/Thesis
Dissertation/thesis number
10148463
ProQuest document ID
1830780524
Copyright
Database copyright ProQuest LLC; ProQuest does not claim copyright in the individual underlying works.
Document URL
https://www.proquest.com/docview/1830780524