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CRISPR/Cas9: An inexpensive, efficie...

Bhattacharya, Dipankan.

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CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus.

Record Type:	Electronic resources : Monograph/item
Title/Author:	CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus./
Author:	Bhattacharya, Dipankan.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
Description:	101 p.
Notes:	Source: Dissertation Abstracts International, Volume: 79-11(E), Section: B.
Contained By:	Dissertation Abstracts International79-11B(E).
Subject:	Developmental biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10907736
ISBN:	9780438191389

CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus.
Bhattacharya, Dipankan.

CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 101 p.

Source: Dissertation Abstracts International, Volume: 79-11(E), Section: B.

Thesis (Ph.D.)--Yale University, 2018.

Congenital malformations are the major cause of infant mortality in the US and Europe. Due to rapid advances in human genomics, we can now efficiently identify sequence variants that may cause disease in these patients. However, establishing disease causality remains a challenge. Additionally, in the case of congenital heart disease, many of the identified candidate genes are either novel to embryonic development or have no known function. Therefore, there is a pressing need to develop inexpensive and efficient technologies to screen these candidate genes for disease phenocopy in model systems and to perform functional studies to uncover their role in development. For this purpose, we sought to test FO CRISPR based gene editing as a loss of function strategy for disease phenocopy in the frog model organism, Xenopus tropicalis . We demonstrate that the CRISPR/ Cas9 system can efficiently modify both alleles in the F0 generation within a few hours post fertilization, recapitulating even early disease phenotypes that are highly similar to knockdowns from morpholino oligos (MOs) in nearly all cases tested. We find that injecting Cas9 protein is dramatically more efficacious and less toxic than cas9 mRNA. We conclude that CRISPR based FO gene modification in X. tropicalis is efficient and cost effective and readily recapitulates disease and MO phenotypes.

ISBN: 9780438191389Subjects--Topical Terms:

592588
Developmental biology.

CRISPR/Cas9: An inexpensive, efficient loss of function tool to screen human disease genes in Xenopus.
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