CRISPR-Cas 9 Mediated Disruption of the FOSL1a Gene in D. rerio

Mentor 1

Ava Udvadia

Start Date

10-5-2022 10:00 AM

Description

Central nervous system regeneration, specifically regeneration of the optic nerve, is not observed in mammals. This phenomenon is observed in D. rerio due to the differential expression of genes in the retinal ganglion cells that allow injured axons to regenerate and restore vision. The goal of the Udvadia Lab is to determine a program of gene expression that regulates successful optic nerve regeneration in zebrafish. The expression of these genes will be compared to the mammalian counterparts to determine which genes can be induced to replicate axon regeneration that is seen in zebrafish. This paper will take this goal one step further in a cross-species analysis between X. laevis and A. mexicanum. Of the genes studied in the Udvadia Lab, JUN is the transcription factor most tightly correlated with the regulation of regenerative gene expression. JUN does not act alone. A repertoire of identified genes included the transcription factor FOSL1. FOSL1 is a protein encoding gene that belongs to the basic leucine zipper transcription factor family. Leucine zipper proteins, encoded by FOSL1, dimerize with proteins from the JUN family. This heterodimer forms the transcription factor complex AP-1, which promotes transcription. Previous research in the Udvadia Lab identified the peak expression of FOSL1 at two days post injury in an adult zebrafish. qPCR techniques will be used to determine levels of gene expression of FOSL1 across the three species mentioned, using RNA extractions from the retinas of the animals. Comparing the expression levels at different post-injury time points will enable us to determine if the temporal expression pattern of FOSL1 is conserved across regenerative species. FOSL1 is not upregulated in response to optic nerve damage in mammalian species, therefore, conserved temporal expression patterns across regenerative species would suggest an important role in regulating gene expression that accompanies successful optic nerve regeneration.

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May 10th, 10:00 AM

CRISPR-Cas 9 Mediated Disruption of the FOSL1a Gene in D. rerio

Central nervous system regeneration, specifically regeneration of the optic nerve, is not observed in mammals. This phenomenon is observed in D. rerio due to the differential expression of genes in the retinal ganglion cells that allow injured axons to regenerate and restore vision. The goal of the Udvadia Lab is to determine a program of gene expression that regulates successful optic nerve regeneration in zebrafish. The expression of these genes will be compared to the mammalian counterparts to determine which genes can be induced to replicate axon regeneration that is seen in zebrafish. This paper will take this goal one step further in a cross-species analysis between X. laevis and A. mexicanum. Of the genes studied in the Udvadia Lab, JUN is the transcription factor most tightly correlated with the regulation of regenerative gene expression. JUN does not act alone. A repertoire of identified genes included the transcription factor FOSL1. FOSL1 is a protein encoding gene that belongs to the basic leucine zipper transcription factor family. Leucine zipper proteins, encoded by FOSL1, dimerize with proteins from the JUN family. This heterodimer forms the transcription factor complex AP-1, which promotes transcription. Previous research in the Udvadia Lab identified the peak expression of FOSL1 at two days post injury in an adult zebrafish. qPCR techniques will be used to determine levels of gene expression of FOSL1 across the three species mentioned, using RNA extractions from the retinas of the animals. Comparing the expression levels at different post-injury time points will enable us to determine if the temporal expression pattern of FOSL1 is conserved across regenerative species. FOSL1 is not upregulated in response to optic nerve damage in mammalian species, therefore, conserved temporal expression patterns across regenerative species would suggest an important role in regulating gene expression that accompanies successful optic nerve regeneration.