alexa Cflarb Complemented the Function of Cflara to Allow Cflara Knock out Zebrafish To Normal Development | OMICS International | Abstract
ISSN: 2168-9296

Cell & Developmental Biology
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Research Article

Cflarb Complemented the Function of Cflara to Allow Cflara Knock out Zebrafish To Normal Development

Huh SJ1, Hwang KS2, Koppula S1, Kim CG3, Kim CH2and Kim CG1*

1Department of Biotechnology, Konkuk University, Chungju, Republic of Korea

2Department of Biology, Chungnam National University, Daejeon, Republic of Korea

3Department of Life Science and Research Institute for Natural Sciences, College of Natural Sciences, Hanyang University, Republic of Korea

4†These authors contributed equally to this work.

*Corresponding Author:
Gil Kim
Department of Biotechnology
Konkuk University, Chungju
Republic of Korea
Tel.: +82-43-840-3614
Fax: +82-43- 846-3616
E-mail: [email protected]

Received date: January 19, 2017; Accepted date: February 22, 2017; Published date: February 28, 2017

Citation: Huh SJ, Hwang KS, Koppula S, Kim CG, Kim CH, et al. (2017) Cflarb Complemented the Function of Cflara to Allow Cflara Knock out Zebrafish to Normal Development. Cell Dev Biol 6:180. doi:10.4172/2168-9296.1000180

Copyright: © 2017 Huh SJ, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Cellular FLICE-inhibitory protein (cFLIP, cflara) is a regulator of death receptor (DR)-induced apoptosis and NF-κB activation. cFLIP is known to prevent activation of the caspase cascade by binding to FADD/caspase-8. Up-regulated cFLIP has been identified in many tumor types, and therefore restoring apoptosis by silencing cFLIP may be one of the more potent strategies in cancer therapeutics. The zebrafish cFLIP gene, cflara, has 2 death effector domains (DEDs) and a single caspase-like domain. Expression of cflara was detected in the zebrafish embryo by RT-PCR and whole-mount in situ hybridization. To study the in vivo function of cflara, we generated a cflara knockout mutant zebrafish using transcription activator-like effector nucleases (TALENs). Frame shift mutation is caused by a 10-bp deletion in the first DED domain. By inbreeding the F1 generation, a homozygous mutant fish was produced and confirmed by PCR. Knockout of cflara leads to abnormal heart development and embryonic lethality in mice. However, mutant zebrafish did not show any differences from wild type in heartbeat rate, survival rate or development. Zebrafish have analogues of both cflara and cflarb. Quantitative PCR showed that cflarb mRNA levels of mutant zebrafish were higher than those in the wild type. In a chemical exposure experiment, mutant zebrafish larvae showed a longer survival rate compared with wild type after CoCl2 treatment. However, no significant difference was observed from cisplatin treatment. This data suggests that cflarb may contribute to normal development and causes a difference in chemical resistance.

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