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B6;129-Rai1tm1Luo Tg(Nr5a1-cre)7Lowl

Summary Data Summary
Official Name B6;129-Rai1tm1Luo Tg(Nr5a1-cre)7Lowl
Common Name B6;129-Rai1tm1Luo Tg(SF1-cre)7Lowl
Description The targeting construct for production of the conditional
Rai1 allele was generated using conventional cloning
approaches with a FRT5-pSV40-Neo-pA-FRT5 cassette and
homology arms amplified by high-fidelity PCR (Phusion,
Finnzymes) from 129X1/SvJ genomic DNA (Jackson Labs). Growth
of the full targeting construct in bacteria at high copy
number led to loss of the DNA segment between loxP sites,
presumably due to the actions of endogenous bacterial
recombinases. To circumvent this issue, the construct was
subcloned into a bacterial artificial chromosome (BAC)
vector prior to the final cloning steps, and the construct
was maintained at 1-2 copy numbers per bacterial cell. After
verification by sequencing, the construct was electroporated
into R1 129Sv/SvJ ES cells, and correctly targeted clones
were identified by long-range PCR (LA Taq, TaKaRa) and DNA
sequencing. Targeted ES cells were microinjected into BL/6
blastocysts, and chimeras were mated to a germline-active
GFP-FlpO transgenic line to remove the neomycin resistance
cassette. Pups lacking the Neo cassette were identified by
PCR and were used to expand the colony. ES cell
manipulations and blastocyst injections were performed by
the Stanford Transgenic Research Facility.

Cre mediated recombination is detected in steroidogenic
factor-1 (SF1)-positive neurons in the ventromedial
hypothalamic nucleus as well as pituitary, gonad, and
adrenal tissue. This transgene includes cre recombinase
under the control of Nr5a1 regulatory sequences.
Development Status Phenotyping ongoing
Creation Method knockin
Investigators 1
Genomics - Modifications 1
Genomics - Transgenes 1
Experiments 2
Publications 1

Liqun LuoStanford University

Genomic Information
GeneAllele 1Allele 2Protocol
Rai1knockin (lox)knockin (lox)Not Specified
Not SpecifiedNr5a1CRENot Specified

PublicationAltmetricsSubmitted ByPubMed IDStatus
Molecular and Neural Functions of Rai1, the Causal Gene for Smith-Magenis Syndrome.
Huang WH, Guenthner CJ, Xu J, Nguyen T, Schwarz LA, Wilkinson AW, Gozani O, Chang HY, Shamloo M, Luo L
Neuron, 2016 (92), 392 - 406
Submitted Externally


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