Zebrafish models

Zebrafish are increasingly being used to study neurodevelopmental disorders, including autism spectrum disorder (ASD). Zebrafish are genetically tractable, transparent during development, and amenable to high-throughput phenotyping, making them a valuable experimental platform to interrogate the biological mechanisms by which genetic mutations affect phenotype and to identity potential therapeutic targets.

Use of mutant zebrafish to study gene function, however, has proven challenging. One reason is that genetic manipulations aimed at producing gene loss-of-function may still leave significant amounts of mRNA produced by the targeted gene, indicating that loss of function of the targeted gene may not have been entirely achieved¹.

To address this issue and facilitate ASD research using zebrafish, SFARI is now curating zebrafish lines with mutations in zebrafish paralogs of ASD risk genes in which gene loss-of-function is validated by directly measuring mRNA or protein levels (where antibodies are available) rather than by assessing phenotype. These include previously existing zebrafish lines, as well as lines that have been deposited and/or validated upon SFARI request, and lines that have been created and validated de novo with SFARI support.

All lines curated by SFARI are listed below and can be found at the Zebrafish International Research Center (ZIRC). Interested investigators should contact ZIRC directly to obtain access.

Available zebrafish models

ARID1B

Zebrafish gene: arid1b

ZIRC allele: y607
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

CHD8

Zebrafish gene: chd8

ZIRC allele: y608
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

DYNC1H1

Zebrafish gene: dync1h1

ZIRC allele: mw20
Created by: Brian Link (Medical College of Wisconsin)²
Method: Chemical mutagenesis
Validation: Reverse transcription polymerase chain reaction (RT-PCR)

DYRK1A

Zebrafish gene: dyrk1aa

ZIRC allele: y656
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

Zebrafish gene: dyrk1ab

ZIRC allele: y657
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

FMR1

Zebrafish gene: fmr1

ZIRC allele: hu2787
Created by: René F. Ketting (University Medical Center Utrecht)³
Method: N-ethyl-N-nitrosourea (ENU) mutagenesis
Validation: Western blot, immunocytochemistry

Zebrafish gene: fmr1

ZIRC allele: hu2898
Created/described by: René F. Ketting, Ph.D. (University Medical Center Utrecht)³
Method: ENU mutagenesis
Validation: Western blot, immunocytochemistry

GRIN2B

Zebrafish gene: grin2ba

ZIRC allele: y658
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

Zebrafish gene: grin2bb

ZIRC allele: y660
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

MECP2

Zebrafish gene: mecp2

ZIRC allele: fh232
Created/described by: Cecilia Moens, Ph.D. (Fred Hutchinson Cancer Research Center)⁴
Method: ENU mutagenesis
Validation: Western blot

MEF2C

Zebrafish gene: mef2ca

ZIRC allele: b1086
Created/described by: Charles Kimmel (Univerity of Oregon)⁵
Method: ENU mutagenesis
Validation: Immunohistochemistry

Zebrafish gene: mef2cb

ZIRC allele: fh288
Created/described by: Cecelia Moens (Fred Hutchinson Cancer Research Center)⁶
Method: ENU mutagenesis
Validation: Immunohistochemistry

PTEN

Zebrafish gene: ptena

ZIRC allele: hu1864
Created by: Jeroen den Hertog, Ph.D. (Leiden University)⁷
Method: ENU mutagenesis
Validation: Western blot

Zebrafish gene: ptenb

ZIRC allele: hu1435
Created by: Jeroen den Hertog, Ph.D. (Leiden University)⁷
Method: ENU mutagenesis
Validation: Western blot

SHANK3

Zebrafish gene: shank3a

ZIRC allele: umi101
Created by: Julia Dallman, Ph.D. (University of Miami)⁹
Method: CRISPR
Validation: Western blot

Zebrafish gene: shank3b

ZIRC allele: umi102
Created by: Julia Dallman, Ph.D. (University of Miami)⁹
Method: CRISPR
Validation: Western blot

SCN1A/SCN2A

Zebrafish gene: scn1lab

ZIRC allele: y609
Created by: Harold Burgess (National Institute of Child Health and Human Development)
Method: CRISPR
Validation: Quantitative polymerase chain reaction (QPCR)

Future zebrafish models

The following lines are expected to be available in 2023:

CNTNAP2
NRXN1
SYNGAP1

For more information about these lines, and general inquiries about SFARI’s experimental models, please contact: [email protected].

References

Anderson J.L. et al. PLoS Genet. 13, e1007105 (2017). PubMed
Insinna C. et al. Neural Dev. 5, 12 (2010). PubMed
den Broeder M.J. et al. PLoS One 4, e7910 (2009) PubMed
Clark K.J. et al. Nat Methods. 8, 6 (2011). Pubmed
Pietri T. et al. Front. Neural Circuits 7, 118 (2013) PubMed
Miller C.T. et al. Developmental Biology 308, 1 (2007). PubMed
Hinits Y.et al. Developmental Biology 369, 2 (2012). Pubmed
Stumpf M. and den Hertog J. PLoS One 11, e0148508 (2016) PubMed
James D.M. et al. Mol. Autism 10, 3 (2019). PubMed