C(Cg)-Tmprss2^{em2.1(TMPRSS2)Synbl}/J

Stock number: 037078
Product name: hTMPRSS2-BALB/c , TMPRSS2-C
Research areas: Cancer Research, Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Mouse/Human Gene Homologs, Research Tools, Virology Research

Description

Human TMPRSS2 (transmembrane protease, serine 2) expression replaces that of the endogenous mouse Tmprss2 gene in these CRISPR/Cas9-generated hTMPRSS2-BALB/c knock-in/knock-out mice that are useful in studies of viral infection and cancer.

Detailed description

TMPRSS2 is a membrane-associated serine protease, expressed in the epithelium of the human respiratory tract and prostate glands. It has been shown to activate hemagglutinin proteins of human influenza viruses and is implicated in prostate carcinogenesis. SARS-CoV-2 binds to the extracellular receptor angiotensin-converting enzyme 2 (ACE2) to enter the host cells and uses the cellular transmembrane serine protease TMPRSS2 as a cooperating partner for S protein cleavage and priming.

Human TMPRSS2 (transmembrane protease, serine 2) expression replaces that of the endogenous mouse Tmprss2 gene in these CRISPR/Cas9-generated hTMPRSS2-BALB/c knock-in/knock-out mice that are useful in studies of viral infection and cancer. SARS-CoV-2 binds to the extracellular receptor angiotensin-converting enzyme 2 (ACE2) to enter the host cells and uses the cellular transmembrane serine protease TMPRSS2 as a cooperating partner for S protein cleavage and priming.

This strain has not yet been published (April, 2022). Please see the reference section below for any updates and further characterization information.

Please note, 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.

Development

Cas9 and gRNA Tmprss2-crRNA_5 (protospacer sequence 5’-GTGCTCAGACTGATACCCG-3’) were used to introduce the following at codon 19 in exon 3 of the mouse Tmprss2 gene on chromosome 16:

• GGSG linker: coding sequence for GGSG linker in-frame with upstream mouse Tmprss2 coding sequence.
• P2A: porcine teschovirus 2A self-cleaving peptide sequence.
• hTMPRSS2 CDS: human TMPRSS2 cDNA coding sequence with SV40 intron inserted for enhanced expression.
• hTMPRSS2 3’UTR: 1837 bp human TMPRSS2 3’UTR.
• hTMPRSS2 3’: 112 bp human intergenic sequence immediately 3’ of TMPRSS2 polyadenylation region.
• Tmprss2-gRNA5 Cassette: “Active genetics” cassette comprised of a mouse U6 promoter-driven Tmprss2-gRNA5 guide RNA with 2xPolIII terminator, all in reverse orientation relative to Tmprss2 gene.
• SA-SV40 poly(A) signal: splice acceptor and SV40 polyadenylation sequence designed to trap and truncate any mouse Tmprss2 isoforms that may splice past the hTMPRSS2 cassette.
• FRT-PGK-EM7-Neo-bGHpA-FRT: Neomycin resistance gene driven by the mouse PGK promoter and with a bovine growth hormone polyadenylation sequence. The FRT sites allow Flp-mediated removal of the neomycin cassette from targeted embryonic stem cell clones as well as collapse of targeted tandem integration events if required.

This strain was created using BALB/cJ-derived embryonic stem cells. The neomycin cassette was excised from the resultant mice through crosses with BALB/cJ background mice expressing Gt(ROSA)26Sor^tm2(FLP*)Sor. The line was maintained on a BALB/cJ genetic background. The donating laboratory Verified the correct integration and verified the lack of off-target backbone integration.

Upon arrival at The Jackson Laboratory, sperm was cryopreserved. Cryorecovery/live colony development will involve BALB/cJ (Stock No. 000651). (April, 2022)

Allele

Symbol: Tmprss2^{em2.1(TMPRSS2)Synbl}
Name: endonuclease-mediated mutation 2.1, Synbal
MGI accession ID: MGI:7261385
Generation method: Endonuclease-mediated
Attributes: Null/Knockout; Inserted expressed sequence; Humanized sequence
Marker symbol: Tmprss2
Marker name: transmembrane protease, serine 2
Marker synonyms: D16Ertd61e; epitheliasin; PRSS10
Chromosome: 16
Strain of origin: BALB/cJ
Molecular note: Cas9 and gRNA Tmprss2-crRNA_5 (protospacer sequence 5’-GTGCTCAGACTGATACCCG-3’) are used to introduce the following sequences at codon 19 in exon 3 of the mouse Tmprss2 gene: GGSG linker, P2A, human TMPRSS2 cDNA coding sequence with SV40, 1837 bp human TMPRSS2 3’UTR, 112 bp human intergenic sequence immediately 3’ of TMPRSS2 polyadenylation region, cassette comprised of a mouse U6 promoter-driven Tmprss2-gRNA5 guide RNA with 2xPolIII terminator, all in reverse orientation relative to Tmprss2 gene, splice acceptor and SV40 polyadenylation sequence designed to trap and truncate any mouse Tmprss2 isoforms that may splice past the hTMPRSS2 cassette. FRT-flanked PGK neomycin cassette with a bGHpA. The FRT sites allow Flp-mediated removal of the neomycin cassette from targeted embryonic stem cell clones as well as collapse of targeted tandem integration events if required. Flp-mediated recombination removed the FRT-flanked neo cassette.