A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene

Chi Hua Chiu; Chris T. Amemiya; Janet L. Carr; Jaya Bhargava; Joseph K. Hwang; Cooduvalli S. Shashikant; Frank H. Ruddle; Günter P. Wagner

doi:10.1007/s004270050016

A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene

Chi Hua Chiu
, Chris T. Amemiya
, Janet L. Carr
, Jaya Bhargava
, Joseph K. Hwang
, Cooduvalli S. Shashikant
, Frank H. Ruddle
, Günter P. Wagner

Obstetrics, Gynecology and Women's Health - Maternal Fetal Medicine

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones.

Original language	English (US)
Pages (from-to)	105-109
Number of pages	5
Journal	Development Genes and Evolution
Volume	210
Issue number	2
DOIs	https://doi.org/10.1007/s004270050016
State	Published - 2000

Bibliographical note

Funding Information:
Acknowledgements We thank Caroline B. Long and Elizabeth Mongillo for technical assistance. C.H.C. is the recipient of an Alfred P. Sloan/National Science Foundation Postdoctoral Research Fellowship in Molecular Evolution. Financial support through National Science Foundation grants IBN-9630567 to G.P.W., F.H.R., C.T.A.; IBN-9614940 to C.T.A.; as well as a National Institutes of Health grant AI 39008-01 to C.T.A. is gratefully appreciated.

Keywords

AbDb-like Hox genes
Cis-regulatory sequences
Homologous recombination
P1/PAC clones
Reporter gene

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10.1007/s004270050016

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Chiu, C. H., Amemiya, C. T., Carr, J. L., Bhargava, J., Hwang, J. K., Shashikant, C. S., Ruddle, F. H., & Wagner, G. P. (2000). A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene. Development Genes and Evolution, 210(2), 105-109. https://doi.org/10.1007/s004270050016

A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene. / Chiu, Chi Hua; Amemiya, Chris T.; Carr, Janet L. et al.
In: Development Genes and Evolution, Vol. 210, No. 2, 2000, p. 105-109.

Research output: Contribution to journal › Article › peer-review

Chiu, CH, Amemiya, CT, Carr, JL, Bhargava, J, Hwang, JK, Shashikant, CS, Ruddle, FH & Wagner, GP 2000, 'A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene', Development Genes and Evolution, vol. 210, no. 2, pp. 105-109. https://doi.org/10.1007/s004270050016

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abstract = "The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones.",

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author = "Chiu, \{Chi Hua\} and Amemiya, \{Chris T.\} and Carr, \{Janet L.\} and Jaya Bhargava and Hwang, \{Joseph K.\} and Shashikant, \{Cooduvalli S.\} and Ruddle, \{Frank H.\} and Wagner, \{G{\"u}nter P.\}",

note = "Funding Information: Acknowledgements We thank Caroline B. Long and Elizabeth Mongillo for technical assistance. C.H.C. is the recipient of an Alfred P. Sloan/National Science Foundation Postdoctoral Research Fellowship in Molecular Evolution. Financial support through National Science Foundation grants IBN-9630567 to G.P.W., F.H.R., C.T.A.; IBN-9614940 to C.T.A.; as well as a National Institutes of Health grant AI 39008-01 to C.T.A. is gratefully appreciated.",

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AU - Amemiya, Chris T.

AU - Carr, Janet L.

AU - Bhargava, Jaya

AU - Hwang, Joseph K.

AU - Shashikant, Cooduvalli S.

AU - Ruddle, Frank H.

AU - Wagner, Günter P.

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PY - 2000

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N2 - The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones.

AB - The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones.

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KW - Homologous recombination

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A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene

Abstract

Bibliographical note

Keywords

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