C57BL/6-Prdm16^em2Snok/J

Stock number: 032162
Product name: Δ47-fPrdm16^-
Research areas: Developmental Biology Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research

Description

Δ47-fPrdm16^- CRISPR/Cas9 generated mutant mice contain a 47 base pair deletion in exon 2 of the PR domain containing 16 (Prdm16) gene, resulting in a premature stop codon. These mice are useful when studying hematopoiesis and inflammatory gene expression in leukemia.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing was used to create a 47 base pair deletion in exon 2 of the PR domain containing 16 (Prdm16) gene, resulting in a premature stop codon. Prdm16 encodes a zinc finger transcriptional coregulator involved in translocations in acute myeloblastic leukemia (AML), myelodysplastic syndromes and T acute lymphoblastic leukemia. PRDM16 is highly expressed in and required for the maintenance of hematopoietic stem cells (HSCs), and can be aberrantly expressed in AML. It is also involved in brown fat, craniofacial, and cardiac development and for the maintenance of subventricular gray zone neural stem cells. This mutation results in absence of the full PRDM16 isoform (fPrdm16) mRNA which uses a promoter in exon 1. Homozygous mice die perinatally, while heterozygous Δ47-fPrdm16^- mice appear normal with slight hematopoietic phenotypes. They show a severe repopulation defect in PB (peripheral blood) and BM, with no defect in homing of donor cells to BM in these mice after transfer. Donor cells from these mice display a stronger and nearly absolute lymphoid bias, and within the lymphoid compartment, a bias towards B-cells, which is not present in Prdm16 KO mice.

Of note, a similar strain with a 13 base pair deletion in exon 2 is available as Stock No. 032161.

Development

Plasmids encoding a signal guide RNAs designed to create a deletion in exon 2 of the PR domain containing 16 (Prdm16) gene and the cas9 nuclease were introduced into the cytoplasm C57BL/6-derived fertilized eggs with well recognized pronuclei. Insertion of guide RNAs resulted in a 47 base pair deletion, creating a premature stop codon. Targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further backcrossed to C57BL/6J mice to develop a congenic colony of Δ47-fPrdm16^- mice. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Prdm16^em2Snok
Name: endonuclease-mediated mutation 2, Hans-Willem Snoeck
MGI accession ID: MGI:6196058
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Synonyms: delta47-fPrdm16^-
Marker symbol: Prdm16
Marker name: PR domain containing 16
Marker synonyms: 5730557K01Rik; CMD1LL; csp1; KMT8F; line 27; LVNC8; MEL1; PFM13
Chromosome: 4
Strain of origin: C57BL/6
Molecular note: CRISPR/cas9 endonuclease mediated genome editing is used to introduce a 47 base pair deletion in exon 2, creating a premature stop codon.