Asset Details
Do you wish to reserve the book?
Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
Do you wish to request the book?
Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Rational targeting of a NuRD subcomplex guided by comprehensive in situ mutagenesis
2019
Overview
Developmental silencing of fetal globins serves as both a paradigm of spatiotemporal gene regulation and an opportunity for therapeutic intervention of β-hemoglobinopathy. The nucleosome remodeling and deacetylase (NuRD) chromatin complex participates in γ-globin repression. We used pooled CRISPR screening to disrupt NuRD protein coding sequences comprehensively in human adult erythroid precursors. Essential for fetal hemoglobin (HbF) control is a non-redundant subcomplex of NuRD protein family paralogs, whose composition we corroborated by affinity chromatography and proximity labeling mass spectrometry proteomics. Mapping top functional guide RNAs identified key protein interfaces where in-frame alleles resulted in loss-of-function due to destabilization or altered function of subunits. We ascertained mutations of
CHD4
that dissociate its requirement for cell fitness from HbF repression in both primary human erythroid precursors and transgenic mice. Finally we demonstrated that sequestering CHD4 from NuRD phenocopied these mutations. These results indicate a generalizable approach to discover protein complex features amenable to rational biochemical targeting.
Comprehensive CRISPR mutagenesis targeting all members of the NuRD complex identifies a specific subcomplex required for fetal globin silencing and informs a rational targeting strategy for elevating globin levels while avoiding cytotoxicity.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 13/109
/ 13/31
/ 38/109
/ 38/22
/ 38/23
/ 38/70
/ 42/41
/ 631/114
/ 631/154
/ 631/1647
/ 631/208
/ 631/61
/ 82/58
/ Animal Genetics and Genomics
/ Animals
/ Biomedical and Life Sciences
/ CRISPR
/ DNA
/ Erythroid Cells - metabolism
/ Fetal Hemoglobin - metabolism
/ Fetuses
/ Fitness
/ Genomes
/ Humans
/ Mi-2 Nucleosome Remodeling and Deacetylase Complex - genetics
/ Mi-2 Nucleosome Remodeling and Deacetylase Complex - metabolism
/ Mice
/ Mutation
/ Protein Interaction Domains and Motifs
/ Proteins
