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A triple polyadenylation stop site was inserted into exon 1, truncating Dlx6os1 splice variant Evf2 (distal-less homeobox 6, opposite strand 1, transcript variant 3 - NR_155496.1) from 3.7kb to 101 bp without affecting the transcription of splice variant Evf1 (distal-less homeobox 6, opposite strand 1, transcript variant 2 - NR_155495.1). RT-PCR confirmed that the expression of Dlx5 and Dlx6 are increased. A floxed neo cassette was removed by cre-mediated recombination.
The Evf2TS targeting construct was generated using lambda phage-based recombineering in E. coli. Following electroporation of W4 embryonic stem cells derived from 129S6/SvEvTac, chimeras were bred to C57BL/6J. Evf2TS(floxed neo)/+ heterozygotes were verified by Southern and crossed to FVB/N-Tg(EIIa-cre)C5379Lmgd/J (RRID:IMSR_JAX:003314) for two generations to remove the selection cassette, followed by additional backcrossing to C57BL/6J to N2 and sib-mating.
Bond AM, Vangompel MJ, Sametsky EA, Clark MF, Savage JC, Disterhoft JF, Kohtz JD. Balanced gene regulation by an embryonic brain ncRNA is critical for adult hippocampal GABA circuitry. Nat Neurosci. 2009 Aug;12(8):1020-7. doi: 10.1038/nn.2371. Epub 2009 Jul 20. (Medline PMID: 19620975)
Cajigas I, Chakraborty A, Swyter KR, Luo H, Bastidas M, Nigro M, Morris ER, Chen S, VanGompel MJW, Leib D, Kohtz SJ, Martina M, Koh S, Ay F, Kohtz JD. The Evf2 Ultraconserved Enhancer lncRNA Functionally and Spatially Organizes Megabase Distant Genes in the Developing Forebrain. Mol Cell. 2018 Sep 20;71(6):956-972.e9. doi: 10.1016/j.molcel.2018.07.024. Epub 2018 Aug 23. (Medline PMID: 30146317)
Cajigas I, Leib DE, Cochrane J, Luo H, Swyter KR, Chen S, Clark BS, Thompson J, Yates JR 3rd, Kingston RE, Kohtz JD. Evf2 lncRNA/BRG1/DLX1 interactions reveal RNA-dependent inhibition of chromatin remodeling. Development. 2015 Aug 1;142(15):2641-52. doi: 10.1242/dev.126318. Epub 2015 Jul 2. (Medline PMID: 26138476)
Berghoff EG, Clark MF, Chen S, Cajigas I, Leib DE, Kohtz JD. Evf2 (Dlx6as) lncRNA regulates ultraconserved enhancer methylation and the differential transcriptional control of adjacent genes. Development. 2013 Nov;140(21):4407-16. doi: 10.1242/dev.099390. Epub 2013 Oct 2. (Medline PMID: 24089468)
Cajigas I, Chakraborty A, Lynam M, Swyter KR, Bastidas M, Collens L, Luo H, Ay F, Kohtz JD. Sox2-Evf2 lncRNA-mediated mechanisms of chromosome topological control in developing forebrain. Development. 2021 Mar 15;148(6):dev197202. doi: 10.1242/dev.197202. (Medline PMID: 33593819)
Colony Surveillance Program and Current Health Reports
Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.
Cryopreserved material may be available upon request, please inquire to mmrrc@missouri.edu for more information.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Sponsored Research.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Sponsored Research
Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.
Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.
1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.
2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.
3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.
4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.