Gene Function, Expression and Regulation in Zebrafish

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications, trials & patents

Abstract

From 8/26/2022 through 8/23/2023 Feldman supported zebrafish research of ten labs, conducted independent research and engaged in institutional service. Work in Precise Genome Editing Independent Research: Testing Reported Improvements in Prime Editing and a New CRISPR Enzyme. May-August 2023. Feldman has completed design, reagent acquisition and initial target locus sequencing for an albino zebrafish rescue scheme to conveniently test repair efficiency of two different albinism-causing mutant alleles. As part of the Sheppard lab projects, Feldman has also planned and acquired preliminary reagents for a strategy to do side-by-side tests of ssODN vs. prime editing-driven single-nucleotide changes. As part of the Farrell lab projects, Feldman has also planned and acquired preliminary reagents to add to his toolkit a new CRISPR enzyme that is reported to cut at distinct PAM-recognition sites than from those currently available. Support of Other Labs (Precise Genome Editing): Sheppard Lab (NICHD): Generation of Four Zebrafish Lines with Single-Nucleotide Precise Genome Edits. May-August 2023. Projects selected, molecular designs completed, target-specific reagents acquired and preliminary target loci sequencing performed. Farrell Lab (NICHD): Generation of Three Zebrafish Lines with 3 Fluorescent Protein Reporter Knockins. May-August 2023. Projects selected, molecular designs completed, target-specific reagents acquired and preliminary target loci sequencing performed. Rogers Lab (NICHD): Generation of one Zebrafish Line with 5 Fluorescent Protein fusion via Knockin. May-August 2023. Project selected, molecular designs completed, target-specific reagents acquired and preliminary target loci sequencing performed. Support of Other Labs (Other Projects): Burgess lab (NICHD): Function of Cacna1c. The Core previously generated a targeted AA-alteration in the cacna1c for which recessive mutants have profound developmental anomalies. This was done for the Golden lab (NIDDK) whose PI (Andy Golden) tragically passed away this year. Characterization of this phenotype in coordination with the Burgess lab (which was also collaborating with the Golden lab) is ongoing. Sackett Lab (NICHD): Assessing Function of Zebrafish Alpha and Beta Tubulin Genes. The degree to which specific tubulin isotypes and/or their post-translational modification are essential for specific aspects of development in any organism remains a surprisingly open question. Feldman trained, and co-supervised two postbac students through this ambitious project to systematically knock-out each zebrafish alpha and beta tubulin isotype in F0 embryos and determine how their absence affects early development. To date, this ongoing project has documented essential embryonic phenotypes for more than five tubulin genes. Dever Lab (NICHD): Translation of Distinct RNA Populations by Eif1 and Eif5. Feldman advised Dever and performed several microinjection experiments to explore differences in phenotypes resulting from ectopic expression of either Eif1 or Eif5 in zebrafish embryos and the possibility that restoration of a balanced Eif1/Eif5 ratio can ameliorate these phenotypes. This project has concluded. Kemper lab (NHLBI): Function of zebrafish rca2.1. The Kemper lab is interested in zebrafish rca2.1s function, because it has certain similarities to human CD46 that are not found in the mouse genome. The Core previously generated two mutant rca2.1 alleles, revealing essential roles in growth and cardiac function. Phenotypic characterization is ongoing. Porter Lab (ex-NICHD): Feldman is finalizing cryopreservation/distribution of lines created/acquired for the Porter lab. Stratakis Lab (ex-NICHD): Feldman is finalizing cryopreservation/distribution of lines created/acquired for the Stratakis lab. Institutional Service: ACUC Membership. Feldman has served on the NICHD ACUC since 2015 and continued in this capacity this year, now (since December 2022) serving as alternate chairperson, meeting monthly to evaluate and decide upon animal-study proposals, renewals and amendments and ad hoc issues relevant to animal welfare.