Kiss1Cre:GFP (v2) knock-in/knock-out mice express cre recombinase fused to a GFP reporter in neurons expressing the Kiss1 (KiSS-1 metastasis-suppressor) gene.
Dr. Richard Palmiter, University of Washington
Transient activation of neurons in the arcuate nucleus of the hypothalamus that express kisspeptin (Kiss1, KiSS-1 metastasis-suppressor) contribute to the occurrence of hot flushes. While the vast majority of patients who suffer from hot flushes are menopausal women, hot flushes can affect both sexes in response to rapid decreases in sex hormone levels, such as that which occurs in men undergoing androgen deprivation therapy for prostate cancer or after castration.
Kiss1Cre:GFP (v2) knock-in/knock-out mice carry a cassette encoding Cre:GFP fusion protein that replaces the coding region of the first Kiss1 coding exon. GFP is expressed constitutively from the Kiss1 promoter. Immunohistochemistry is needed to visualize the fluorescence in this line.
When crossed to a conditional reporter line (Ai14, Stock No. 007914), Kiss1 neurons are faithfully labeled in the rostral periventricular area, bed nucleus of the stria terminalis, arcuate nucleus and medial amygdala, with little to no ectopic expression.
The mutation in this strain represents an improved version of the Kiss1tm1.1(cre/EGFP)Stei allele (Stock No. 017701, also called Kiss1Cre:GFP v1) which is prone to ectopic expression when crossed to conditional reporter mice, in addition to retaining low levels of Kiss1 expression. Unlike the previous allele, homozygotes of the Kiss1Cre:GFP (v2) line are Kiss1 null and infertile. Homozygous Kiss1Cre:GFP (v2) males have diminished testosterone levels compared to heterozygotes.
Barring genetic crosses, the Kiss1Cre:GFP v1 line (Stock No. 017701) remains a useful tool for visualizing Kiss1 neurons for electrophysiological recordings with the GFP-fused cre-reporter transgene. No immunohistochemistry is required to visualize GFP expression in the Kiss1Cre:GFP v1 line. Also, viral injections into adult v1 mice, as opposed genetic crosses, can be used to gain control of those Kiss1 neurons and thereby avoid developmental ectopic recombination.
In brief, a Cre:GFP fusion carrying Myc 3' UTR pA, and an FRT-SvNeo-FRT cassette were introduced to the end of the first Kiss1 exon, such that the sequence between the start codon and the end of that exon is deleted. The mutation was created via homologous recombination in C57BL/6-derived embryonic stem (ES) cells. The FRT-flanked Neo gene was removed by breeding resultant heterozygous mice with animals expressing FLPo recombinase (B6.129S4-Gt(ROSA)26Sortm2(FLP*)Sor, Stock No. 012930). This strain was backcrossed to C57BL/6 for 13 generations by the donating laboratory.
The mutation in this strain represents an improved version of the Kiss1tm1.1(cre/EGFP)Stei allele (Stock No. 017701, also called Kiss1Cre:GFP v1), which was prone to ectopic expression when crossed to conditional reporter mice, as well as retained levels of low expression. The original mutation was modified by: (a) removing the nuclear localization signal from cre, (b) using a non-optimal initiation codon, (c) removing part of the N-terminal sequence of cre, (d) adding a 3' untranslated region from the Myc gene that promotes a short mRNA half-life, (e) deleting part of exon 1 that includes the signal peptide, and (f) performing the gene targeting in C57BL/6 ES cells.
|Allele Name||targeted mutation 1.1, Richard D Palmiter|
|Allele Type||Targeted (Recombinase-expressing, Reporter, Null/Knockout)|
|Allele Synonym(s)||Kiss1Cre:GFP; Kiss1Cre (v2)|
|Gene Symbol and Name||Kiss1, KiSS-1 metastasis-suppressor|
|Strain of Origin||C57BL/6|
|Molecular Note||The targeting construct contains a Cre:GFP fusion carrying Myc 3' UTR pA, and an FRT-SvNeo-FRT cassette introduced to the end of the first exon, such that the sequence between the start codon and the end of that exon are deleted. Flp-mediated recombination removed the FRT-flanked neo cassette. The construct removes the nuclear localization signal from cre, uses a non-optimal initiation codon, removes part of the N-terminal sequence of cre, adds a 3' untranslated region from the Myc gene that promotes a short mRNA half-life, and deletes part of exon 1 that includes the signal peptide.|
Heterozygotes are viable and fertile. Homozygotes are infertile.
When using the KissCre (v2), KissCre:GFP (v2)
mouse strain in a publication, please cite the originating article(s) and include JAX stock #033169 in your Materials and Methods section.
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.