Creation of a Deletion Series of Mouse YACs Covering a 500 Kb Region Around Xist

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1994 May 25

PMID 8208607

Citations 10

Authors

E Heard

P Avner

R Rothstein

Affiliations

Soon will be listed here.

Abstract

Two mouse YACs, PA-2 and PA-3, contain the Xist gene and are 460 kb and 3.3 Mb long respectively. While PA-2 is non-chimeric, PA-3 contains a substantial proportion of non-contiguous DNA. As a prerequisite to functional studies of the role of this region in X inactivation, we have created a deletion series of YACs that are spaced at approximately 50 kb intervals and were able to eliminate the unwanted chimeric sequences in YAC PA-3. For this purpose, we have constructed mouse B1 fragmentation vectors based on those described for human Alu fragmentation. Having created this series of YAC deletion derivatives, we were able to eliminate efficiently the 10-15% aberrant YACs that arise during the course of a fragmentation experiment by assessing their marker content. The overlap and the opposite orientation of the two YAC inserts permitted the creation of deletions on both sides of the 500 kb region around Xist. The use of this series of YACs in a biological assay will help us define the extent of the sequences necessary to bring about X chromosome inactivation.

Citing Articles

The bipartite TAD organization of the X-inactivation center ensures opposing developmental regulation of Tsix and Xist.

van Bemmel J, Galupa R, Gard C, Servant N, Picard C, Davies J Nat Genet. 2019; 51(6):1024-1034.

PMID: 31133748 PMC: 6551226. DOI: 10.1038/s41588-019-0412-0.

Cis- and trans-regulation in X inactivation.

van Bemmel J, Mira-Bontenbal H, Gribnau J Chromosoma. 2015; 125(1):41-50.

PMID: 26198462 PMC: 4761001. DOI: 10.1007/s00412-015-0525-x.

Chromosome integrity in Saccharomyces cerevisiae: the interplay of DNA replication initiation factors, elongation factors, and origins.

Huang D, Koshland D Genes Dev. 2003; 17(14):1741-54.

PMID: 12865298 PMC: 196182. DOI: 10.1101/gad.1089203.

Differential expression of B1-containing transcripts in Leishmania-exposed macrophages.

Ueda Y, Chaudhuri G J Biol Chem. 2000; 275(25):19428-32.

PMID: 10781585 PMC: 3086771. DOI: 10.1074/jbc.M001336200.

Functional analysis of the DXPas34 locus, a 3' regulator of Xist expression.

Debrand E, Chureau C, Arnaud D, Avner P, Heard E Mol Cell Biol. 1999; 19(12):8513-25.

PMID: 10567576 PMC: 84967. DOI: 10.1128/MCB.19.12.8513.

References

Cattanach B, Pollard C, Perez J . Controlling elements in the mouse X-chromosome. I. Interaction with the X-linked genes. Genet Res. 1969; 14(3):223-35. DOI: 10.1017/s0016672300002068. View

Burgers P, Percival K . Transformation of yeast spheroplasts without cell fusion. Anal Biochem. 1987; 163(2):391-7. DOI: 10.1016/0003-2697(87)90240-5. View

Quentin Y . Successive waves of fixation of B1 variants in rodent lineage history. J Mol Evol. 1989; 28(4):299-305. DOI: 10.1007/BF02103425. View

Brownstein B, Silverman G, LITTLE R, Burke D, Korsmeyer S, Schlessinger D . Isolation of single-copy human genes from a library of yeast artificial chromosome clones. Science. 1989; 244(4910):1348-51. DOI: 10.1126/science.2544027. View

Pavan W, Hieter P, Reeves R . Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes. Proc Natl Acad Sci U S A. 1990; 87(4):1300-4. PMC: 53462. DOI: 10.1073/pnas.87.4.1300. View

Pachnis V, Pevny L, Rothstein R, Costantini F . Transfer of a yeast artificial chromosome carrying human DNA from Saccharomyces cerevisiae into mammalian cells. Proc Natl Acad Sci U S A. 1990; 87(13):5109-13. PMC: 54271. DOI: 10.1073/pnas.87.13.5109. View

Brown C, Lafreniere R, Powers V, Sebastio G, Ballabio A, PETTIGREW A . Localization of the X inactivation centre on the human X chromosome in Xq13. Nature. 1991; 349(6304):82-4. DOI: 10.1038/349082a0. View

Borsani G, Tonlorenzi R, Simmler M, Dandolo L, Arnaud D, Capra V . Characterization of a murine gene expressed from the inactive X chromosome. Nature. 1991; 351(6324):325-9. DOI: 10.1038/351325a0. View

Brockdorff N, Ashworth A, Kay G, Cooper P, Smith S, McCabe V . Conservation of position and exclusive expression of mouse Xist from the inactive X chromosome. Nature. 1991; 351(6324):329-31. DOI: 10.1038/351329a0. View

10.

Larin Z, Monaco A, Lehrach H . Yeast artificial chromosome libraries containing large inserts from mouse and human DNA. Proc Natl Acad Sci U S A. 1991; 88(10):4123-7. PMC: 51610. DOI: 10.1073/pnas.88.10.4123. View

11.

Cattanach B, Rasberry C, Evans E, Dandolo L, Simmler M, Avner P . Genetic and molecular evidence of an X-chromosome deletion spanning the tabby (Ta) and testicular feminization (Tfm) loci in the mouse. Cytogenet Cell Genet. 1991; 56(3-4):137-43. DOI: 10.1159/000133070. View

12.

Pavan W, Hieter P, Sears D, Burkhoff A, Reeves R . High-efficiency yeast artificial chromosome fragmentation vectors. Gene. 1991; 106(1):125-7. DOI: 10.1016/0378-1119(91)90576-w. View

13.

Huxley C, Gnirke A . Transfer of yeast artificial chromosomes from yeast to mammalian cells. Bioessays. 1991; 13(10):545-50. DOI: 10.1002/bies.950131009. View

14.

Brockdorff N, Ashworth A, Kay G, McCabe V, Norris D, Cooper P . The product of the mouse Xist gene is a 15 kb inactive X-specific transcript containing no conserved ORF and located in the nucleus. Cell. 1992; 71(3):515-26. DOI: 10.1016/0092-8674(92)90519-i. View

15.

Brown C, Hendrich B, Rupert J, Lafreniere R, Xing Y, Lawrence J . The human XIST gene: analysis of a 17 kb inactive X-specific RNA that contains conserved repeats and is highly localized within the nucleus. Cell. 1992; 71(3):527-42. DOI: 10.1016/0092-8674(92)90520-m. View

16.

Reeves R, Pavan W, Hieter P . Yeast artificial chromosome modification and manipulation. Methods Enzymol. 1992; 216:584-603. DOI: 10.1016/0076-6879(92)16051-k. View

17.

Kay G, Penny G, Patel D, Ashworth A, Brockdorff N, Rastan S . Expression of Xist during mouse development suggests a role in the initiation of X chromosome inactivation. Cell. 1993; 72(2):171-82. DOI: 10.1016/0092-8674(93)90658-d. View

18.

Capecchi M . Mouse genetics. YACs to the rescue. Nature. 1993; 362(6417):205-6. DOI: 10.1038/362205a0. View

19.

Heard E, Simmler M, Larin Z, Rougeulle C, Courtier B, Lehrach H . Physical mapping and YAC contig analysis of the region surrounding Xist on the mouse X chromosome. Genomics. 1993; 15(3):559-69. DOI: 10.1006/geno.1993.1108. View

20.

Gnirke A, Huxley C, Peterson K, Olson M . Microinjection of intact 200- to 500-kb fragments of YAC DNA into mammalian cells. Genomics. 1993; 15(3):659-67. DOI: 10.1006/geno.1993.1121. View