Document Detail


Megabase-scale deletion using CRISPR/Cas9 to generate a fully haploid human cell line.
MedLine Citation:
PMID:  25373145     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Near-haploid human cell lines are instrumental for genetic screens and genome engineering as gene inactivation is greatly facilitated by the absence of a second gene copy. However, no completely haploid human cell line has been described, hampering the genetic accessibility of a subset of genes. The near-haploid human cell line HAP1 contains a single copy of all chromosomes except for a heterozygous 30-megabase fragment of Chromosome 15. This large fragment encompasses 330 genes and is integrated on the long arm of Chromosome 19. Here, we employ a CRISPR/Cas9-based genome engineering strategy to excise this sizeable chromosomal fragment and to efficiently and reproducibly derive clones that retain their haploid state. Importantly, spectral karyotyping and single-nucleotide polymorphism (SNP) genotyping revealed that engineered-HAPloid (eHAP) cells are fully haploid with no gross chromosomal aberrations induced by Cas9. Furthermore, whole-genome sequence and transcriptome analysis of the parental HAP1 and an eHAP cell line showed that transcriptional changes are limited to the excised Chromosome 15 fragment. Together, we demonstrate the feasibility of efficiently engineering megabase deletions with the CRISPR/Cas9 technology and report the first fully haploid human cell line.
Authors:
Patrick Essletzbichler; Tomasz Konopka; Federica Santoro; Doris Chen; Bianca V Gapp; Robert Kralovics; Thijn R Brummelkamp; Sebastian M B Nijman; Tilmann Bürckstümmer
Related Documents :
25193415 - 3p interstitial deletion including prickle2 in identical twins with autistic features.
25232865 - Wt1 interacts with mad2 and regulates mitotic checkpoint function.
94005 - Nucleolar organizer regions (nors) in chinese hamster chromosomes as visualized by coom...
6160765 - A search for protein cores in chromosomes: is the scaffold an artifact?
9914385 - Trichlorfon exposure, spindle aberrations and nondisjunction in mammalian oocytes.
17010155 - Chromosomal loci that influence oral nicotine consumption in c57bl/6j x c3h/hej f2 inte...
Publication Detail:
Type:  Journal Article     Date:  2014-11-04
Journal Detail:
Title:  Genome research     Volume:  24     ISSN:  1549-5469     ISO Abbreviation:  Genome Res.     Publication Date:  2014 Dec 
Date Detail:
Created Date:  2014-12-02     Completed Date:  -     Revised Date:  2014-12-02    
Medline Journal Info:
Nlm Unique ID:  9518021     Medline TA:  Genome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2059-65     Citation Subset:  IM    
Copyright Information:
© 2014 Essletzbichler et al.; Published by Cold Spring Harbor Laboratory Press.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Single haplotype assembly of the human genome from a hydatidiform mole.
Next Document:  Genome accessibility is widely preserved and locally modulated during mitosis.