By: Henry Prewitt
Max Luf is an undergraduate student at Fordham studying integrative neuroscience. Max’s commitments to medical sciences extend to his extracurricular involvements. He is an EMT and officer with Fordham University EMS and the Fordham Integrative Neuroscience Student Association’s secretary. After graduation in 2022, Max plans to enroll in a master’s program or conduct an extended research project before pursuing an M.D.-Ph.D.
Max started doing undergraduate research in the spring of 2020 when he joined Dr. Dubrovsky’s Drosophila genetics lab at Fordham. Max is a collaborator on a broader endeavor in which Dr. Dubrovsky’s lab is studying the gene RNaseZ. This gene is involved in the maturation of tRNA in the common fruit fly species Drosophila melanogaster. What makes it so special is that it is homologous to the human gene ELAC2. RNaseZ and ELAC2 are extremely similar in both function and sequence, which allows the lab to study its importance in humans by observing how it affects flies.
The lab has discovered that cells will die if RNaseZ is absent. Furthermore, the lab has observed from existing research and patient records that people with ELAC2 mutations often develop the heart condition hypertrophic cardiomyopathy. This condition thickens the walls of the heart and reduces its ability to pump blood. Another observed trend in certain patients with ELAC2 mutations is the development of encephalopathies — or diseases of the brain — such as microcephaly (an abnormally small head) and untreatable epilepsy. It is currently unclear how mutations of the ELAC2 gene cause these encephalopathies.
Max’s research will give us a better understanding of the interaction between ELAC2 and the brain. He will observe the effects of various RNaseZ mutations in the brain of Drosophila using the CRISPR-Cas9 system. To isolate the neurological effects of RNaseZ, Max will recreate specific RNaseZ mutations, but only in the brain of flies. This is unlike normal RNaseZ or ELAC2 mutations, which would change the sequence of the DNA in the entire body. Last semester (fall 2020), Max created a bacterial plasmid with the DNA promoter sequence ELAV, which promotes the transcription of Cas9 protein in flies, but only in their brains. Max extracted DNA from Drosophila, isolated the ELAV promoter sequence, amplified it using the polymerase chain reaction, and then used various lab techniques to insert the promoter into bacterial plasmids. He noted that the creation of the plasmid was a significant process in and of itself. Max then had an external lab insert that plasmid into Drosophila embryos. When those flies are born, some of them will have incorporated the ELAV-Cas9 plasmid into their chromosomes.
Max is currently screening these flies to identify which ones have successfully incorporated the plasmid. Once he has identified the flies with the Cas9 protein, he can breed them with flies with a specific type of guide RNA. The offspring from that cross should then wholly lack RNaseZ in their brains. From there, Max can observe the resulting neurological effects. He will then repeat that process, but the guide RNA will be different. Instead of cutting out the RNaseZ gene completely, it will have a sequence that emulates those observed in human patients with ELAC2 mutations who experienced certain encephalopathies. Max’s findings will tell us if the observed encephalopathies in human patients are caused by the mutation of RNaseZ or ELAC2 in brain cells or if they are caused by issues in other parts of the body such as hypertrophic cardiomyopathy. Ultimately, this project is extremely valuable because it will provide crucial information on the role of RNaseZ in the nervous system and has the potential to lead to therapies and treatments for human patients with mutations of the ELAC2 gene.