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The Developmental of a Conditional Knock-In Mouse Model for the Study GEMIN5 Dysfunction

Nelson, Charles H (2023) The Developmental of a Conditional Knock-In Mouse Model for the Study GEMIN5 Dysfunction. Master's Thesis, University of Pittsburgh. (Unpublished)

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Rare pathogenic mutations of the multidomain RBP GEMIN5 have recently been shown to cause NEDCAM, a unique neurodevelopmental disorder characterized by cerebellar volume loss and motor dysfunction. While several models have been used to investigate the pathogenesis of NEDCAM, one more closely related to humans is required for future research. As a rare disease, NEDCAM does not pose a threat to a majority of people, however, rare diseases have a high societal cost, a relevant public health concern. As Gemin5 pathogenic mutations are embryonically lethal in mice, a conditional model was needed to facilitate investigation. Here, I describe the creation of a conditional mouse model for the study of the rare pathogenic Gemin5 variant p.L1067P. Using Gemin5 floxed mice and mice ubiquitously expressing Cre-ERT2, a three-step breeding scheme generated mice with temporally controlled homozygous expression of Gemin5L1067P/L1067P at expected Mendelian ratios. After five days of daily 100 mg/kg tamoxifen injections starting at 3.5 weeks of age, Gemin5L1067P/L1067P Flox/UBC-Cre mice showed significant differences in bodyweight, body length, fore and hindlimb stride length, but not cerebellar volume when compared to control mice lacking Cre expression. This result was also observed in mice receiving five days of daily 50 mg/kg tamoxifen injections at 3 weeks of age with the addition of significantly different forelimb-hindlimb step overlay, a measure of coordination. Open field test results also showed greater time spent at rest, less time moving slow and fast, shorter distance traveled, and lower average and maximum velocities when compared with controls. Despite this, no significant cerebellar volume difference was detected when compared with controls. Overall, these results suggest that Gemin5L1067P/L1067P Flox/UBC-Cre mice display phenotypic markers with similarity to some aspects of NEDCAM in humans. Further, the lack of cerebellar volume loss in these mice provides evidence that cerebellar hypoplasia, rather than atrophy, may be the cause of the volume loss observed in human patients. Nevertheless, the model’s temporal control allows for future studies to be conducted at earlier time points, allowing the model to be further refined.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Nelson, Charles Hchn63@pitt.educhn630000-0002-2463-2155
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairPandey, Udai B.udai@pitt.eduudai
Thesis AdvisorPandey, Udai B.udai@pitt.eduudai
Committee MemberDemirci, F. Yesimfyd1@pitt.edufyd1
Committee MemberKemaladewi,
Committee MemberUrban, Zsolturbanz@pitt.eduurbanz
Date: 11 May 2023
Date Type: Publication
Defense Date: 26 April 2023
Approval Date: 11 May 2023
Submission Date: 27 April 2023
Access Restriction: 2 year -- Restrict access to University of Pittsburgh for a period of 2 years.
Number of Pages: 78
Institution: University of Pittsburgh
Schools and Programs: School of Public Health > Human Genetics
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: GEMIN5, Cre-loxP, Mouse, Conditional model, NEDCAM, SMN Complex
Date Deposited: 11 May 2023 16:14
Last Modified: 11 May 2023 16:14


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