| Autor: |
Lou WP; School of Life Sciences, South China Normal University; The Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC)., Wang L; Institute for Brain Research and Rehabilitation (IBRR), South China Normal University., Long C; School of Life Sciences, South China Normal University., Liu L; Institute for Brain Research and Rehabilitation (IBRR), South China Normal University; liulei@scnu.edu.cn., Fei JF; Institute for Brain Research and Rehabilitation (IBRR), South China Normal University; jifengfei@m.scnu.edu.cn. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2019 Jul 09 (149). Date of Electronic Publication: 2019 Jul 09. |
| DOI: |
10.3791/59850 |
| Abstrakt: |
The axolotl has the unique ability to fully regenerate its spinal cord. This is largely due to the ependymal cells remaining as neural stem cells (NSCs) throughout life, which proliferate to reform the ependymal tube and differentiate into lost neurons after spinal cord injury. Deciphering how these NSCs retain pluripotency post-development and proliferate upon spinal cord injury to reform the exact pre-injury structure can provide valuable insight into how mammalian spinal cords may regenerate as well as potential treatment options. Performing gene knock-outs in specific subsets of NSCs within a restricted time period will allow study of the molecular mechanisms behind these regenerative processes, without being confounded by development perturbing effects. Described here is a method to perform gene knock-out in axolotl spinal cord NSCs using the CRISPR-Cas9 system. By injecting the CAS9-gRNA complex into the spinal cord central canal followed by electroporation, target genes are knocked out in NSCs within specific regions of the spinal cord at a desired timepoint, allowing for molecular studies of spinal cord NSCs during regeneration. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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