CRISPR/Cas9-mediated correction of MITF homozygous point mutation in a Waardenburg syndrome 2A pig model.

Molecular Therapy. Nucleic Acids
Jing YaoJianguo Zhao

Abstract

Gene therapy for curing congenital human diseases is promising, but the feasibility and safety need to be further evaluated. In this study, based on a pig model that carries the c.740T>C (L247S) mutation in MITF with an inheritance pattern and clinical pathology that mimics Waardenburg syndrome 2A (WS2A), we corrected the point mutation by the CRISPR-Cas9 system in the mutant fibroblast cells using single-stranded oligodeoxynucleotide (ssODN) and long donor plasmid DNA as the repair template. By using long donor DNA, precise correction of this point mutation was achieved. The corrected cells were then used as the donor cell for somatic cell nuclear transfer (SCNT) to produce piglets, which exhibited a successfully rescued phenotype of WS2A, including anophthalmia and hearing loss. Furthermore, engineered base editors (BEs) were exploited to make the correction in mutant porcine fibroblast cells and early embryos. The correction efficiency was greatly improved, whereas substantial off-targeting mutations were detected, raising a safety concern for their potential applications in gene therapy. Thus, we explored the possibility of precise correction of WS2A-causing gene mutation by the CRISPR-Cas9 system in a large-animal model, s...Continue Reading

References

Nov 1, 1995·Human Molecular Genetics·M TassabehjiW Reardon
Feb 15, 2003·Japanese Journal of Ophthalmology·Naonori OhnoManabu Mochizuki
Aug 2, 2007·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Amit KochharRichard J H Smith
May 20, 2009·Bioinformatics·Heng Li, Richard Durbin
Jan 5, 2013·Science·Le CongFeng Zhang
Oct 26, 2013·Nature Protocols·F Ann RanFeng Zhang
Feb 14, 2014·PloS One·Verónica Ponce de LeónEdith Hummler
Mar 4, 2014·Nature Biotechnology·Jeffry D Sander, J Keith Joung
Apr 1, 2014·Nature Biotechnology·Hao YinDaniel G Anderson
Apr 4, 2014·Bioinformatics·Anthony M BolgerBjoern Usadel
Jun 24, 2014·Scientific Reports·Masafumi InuiShuji Takada
Nov 15, 2014·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Weiwei GuoShi-Ming Yang
Dec 17, 2014·Documenta Ophthalmologica. Advances in Ophthalmology·Daphne L McCullochMichael Bach
Jun 24, 2015·Clinical Genetics·J SongI J Dhooge
Nov 20, 2015·Nucleic Acids Research·Melissa J LandrumDonna R Maglott
Jan 29, 2016·Nature Communications·Jun SongJifeng Zhang
Jun 9, 2016·Genome Biology·William McLarenFiona Cunningham
Jul 14, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Megan D HobanDonald B Kohn
Nov 28, 2016·American Journal of Human Genetics·Aman GeorgeBrian P Brooks
Sep 25, 2017·Protein & Cell·Puping LiangJunjiu Huang

❮ Previous
Next ❯

Software Mentioned

LoFreq
Strelka2
Burrows
Hi
EditR
Cas
TOM
OFFinder
Mutect2
GATK

Related Concepts

Related Feeds

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

Auditory Perception

Auditory perception is the ability to receive and interpret information attained by the ears. Here is the latest research on factors and underlying mechanisms that influence auditory perception.

CRISPR Genome Editing & Therapy

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on the application of this system for gene editing and therapy in human diseases.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

© 2021 Meta ULC. All rights reserved