C57BL/6J-Tg(CAG-ACE2)GT5-027Cyt/J

Stock number: 034900
Product name: CAG-hACE2
Research areas: Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Mouse/Human Gene Homologs, Research Tools, Virology Research

Description

CAG-hACE2 transgenic mice express human ACE2, the receptor used by severe acute respiratory syndrome coronavirus (SARS-CoV) to gain cellular entry. Since the CAG promoter is highly active in a variety of tissues, this strain is useful for monitoring severe COVID-19 and sex-biased responses to SARS-CoV-2 infection, and may aid in the development of vaccines and therapeutic treatments for the disease.

These mice should be handled in a manner consistent with CDC/ABSA/WHO guidelines for prevention of human infection with the SARS-CoV-2 virus. Proper PPE and handling methods should be used at all times when working with these mice. Additional important guidelines for using SARS-CoV-2 susceptible mouse lines.

Detailed description

CAG-hACE2 transgenic mice express human ACE2, the receptor used by severe acute respiratory syndrome coronavirus (SARS-CoV) to gain cellular entry under control of the CAG promoter in a variety of tissues. The CAG-hACE2 transgenic mice display increased expression of hACE2 transgene in lung, brain and kidney and relatively lower expression in duodenum, heart and liver. Homozygous mice are viable, but are smaller than hemizygous mice by P10-P12, and exhibit postnatal lethality soon after. Hemizygous males and females are viable and fertile and have equal transgene expression levels. Hemizygous mice are highly susceptibility to SARS-CoV-2 infection, with 100 PFU of SARS-CoV-2 being sufficient to induce body weight loss and 87.5% mortality at 9 days post-infection with a sole (female) survivor. Mortality is 100% at a higher titer of 1000 PFU. At lower viral titers, female mice exposed to SARS-CoV-2 infection suffer much less weight loss than male mice, implying sex-biased responses to SARS-CoV-2 infection. Expression of SARS-CoV-2 Spike protein in neuronal cultures from these mice results in altered synaptic responses of cultured neurons.

Please note, these CAG-hACE2 mice should be handled in a manner consistent with CDC/ABSA/WHO guidelines for prevention of human infection with the SARS-CoV-2 virus. Proper PPE and handling methods should be used at all times when working with these mice. Additional important guidelines for using SARS-CoV-2 susceptible mouse lines.

Development

The CAG-hACE2 transgene has, from 5’ to 3’, 2 copies of a chicken HS4 insulator sequence, a CMV enhancer/chicken beta-actin (CAG) promoter, full length a hemaglutatin (HA) tagged human angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 (hACE2) cDNA, and 2 more copies of the chicken HS4 insulator. This transgene was microinjected into fertilized C57BL/6J oocytes. CAG-hACE2 mice from founder line GT5-027, with 2 copies of the transgene, were subsequently maintained by breeding transgenic mice with C57BL/6J mice for at least 3 generations. Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To generate our live colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J mice (Stock No. 000664).

Allele

Symbol: Tg(CAG-ACE2)GT5-027Cyt
Name: transgene insertion GT5-027, Ching-Yen Tsai
MGI accession ID: MGI:6711183
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence; Epitope tag
Synonyms: CAG-hACE2
Marker symbol: Tg(CAG-ACE2)GT5-027Cyt
Marker name: transgene insertion GT5-027, Ching-Yen Tsai
Chromosome: UN
Strain of origin: C57BL/6J
Molecular note: The transgenic construct contains 2 copies of a chicken HS4 insulator sequence, a CMV enhancer/chicken beta-actin (CAG) promoter, full length a hemaglutatin (HA) tagged human angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 (hACE2) cDNA, and 2 more copies of the chicken HS4 insulator. Founder lines GT5-027 and GT4-008 were generated. GT5-027 carries two copies of the transgene and was used in experiments. High expression levels are detected in lung, kidney and brain.