Establishment and characterisation of CRTC1 knockout in rodent hippocampal neurons

CRTC1 is a transcriptional modulator that is crucial for learning and memory. At present, a conditional knockout mouse has never been studied either in neuronal cultures or live animals. In this report, we propose the use of a single lentiviral vector to deliver Cas9 and guide RNAs (gRNAs) into neur...

Full description

Saved in:
Bibliographic Details
Main Author: Koh, Tyler Bo Kai
Other Authors: Ch'ng Toh Hean
Format: Final Year Project
Language:English
Published: 2017
Subjects:
Online Access:http://hdl.handle.net/10356/71555
Tags: Add Tag
No Tags, Be the first to tag this record!
Institution: Nanyang Technological University
Language: English
Description
Summary:CRTC1 is a transcriptional modulator that is crucial for learning and memory. At present, a conditional knockout mouse has never been studied either in neuronal cultures or live animals. In this report, we propose the use of a single lentiviral vector to deliver Cas9 and guide RNAs (gRNAs) into neurons to knockout the expression of CRTC1 in hippocampal neurons in vitro and in vivo. Our lentiviral vectors efficiently knockouts CRTC1 expression in primary hippocampal cultures while results are pending for in vivo microinjections of the virus. A preliminary study on the effects of neurons lacking CRTC1 expression indicate an increase in spine area and length while the distribution and density of spines remain unaltered.