Creatine Transporter Deficiency examined using QRT-PCR of SLC6A8A and SLC6A8B knockout zebrafish embryos
Katherine Bassler, Joseph Sheheen, Dr. Susan C. Chapman
We have successfully performed qRT-PCR for seven genes (dlx2a, eomesa, lhx6, lim3, slc6a8a, slc6a8b, and zash1a) that have roles in patterning the developing zebrafish brain. Two control genes, actb and gapdh, were also successfully tested.
We have shown that we can harvest zebrafish tissues, make RNA and cDNA for use in qRT-PCR analysis.
We have completed harvesting head tissues from SLC6A8B knockout embryos and AB wildtype controls. SLC6A8A knockout embryos have been identified by genotyping and are being bred to provide tissues for RNA extraction. Once all tissues have been harvested and cDNA prepared, we will perform qRT-PCR analysis between the knockout embryos and control embryos to determine the effect of SLC6A8 mutations on zebrafish brain development.
Using zebrafish, we aim to determine which candidate patterning genes are affected during brain patterning when the creatine transporter is mutated. This will lead to a greater understanding of the effects of SLC6A8 mutations on human health outcomes.
- Mercimek-Mahmutoglu, S. Creatine Deficiency Syndromes https://www.ncbi.nlm.nih.gov/books/NBK3794/ (accessed Aug 18, 2019).
- SLC6A8 gene – Genetics Home Reference https://ghr.nlm.nih.gov/gene/SLC6A8 (accessed Aug 18, 2019).