Development of a Mouse Model of Hematopoietic Loss of Y Chromosome.
Bone marrow reconstitution
Cas9-knockin mice
Clonal hematopoiesis
Heart failure
Hematopoietic mosaic loss of Y chromosome (mLOY)
gRNA-encoding lentivirus
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Aug 2023
05 Aug 2023
Historique:
received:
25
12
2022
revised:
27
02
2023
accepted:
08
05
2023
medline:
14
8
2023
pubmed:
14
8
2023
entrez:
14
8
2023
Statut:
epublish
Résumé
This protocol describes the generation of chimeric mice in which the Y chromosome is deleted from a proportion of blood cells. This model recapitulates the phenomenon of hematopoietic mosaic loss of Y chromosome (mLOY), which is frequently observed in the blood of aged men. To construct mice with hematopoietic Y chromosome loss, lineage-negative cells are isolated from the bone marrow of ROSA26-Cas9 knock-in mice. These cells are transduced with a lentivirus vector encoding a guide RNA (gRNA) that targets multiple repeats of the Y chromosome centromere, effectively removing the Y chromosome. These cells are then transplanted into lethally irradiated wildtype C57BL6 mice. Control gRNAs are designed to target either no specific region or the fourth intron of
Identifiants
pubmed: 37575386
doi: 10.21769/BioProtoc.4729
pii: e4729
pmc: PMC10415196
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4729Informations de copyright
©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no competing interests.
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