Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites

E.R. Coppoolse, M.J. de Vroomen, F. van Gennip, B.J.M. Hersmus, M.J. van Haaren

Research output: Contribution to journalArticleAcademicpeer-review

16 Citations (Scopus)

Abstract

Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable.
Original languageEnglish
Pages (from-to)687-698
JournalPlant Molecular Biology
Volume58
Issue number5
DOIs
Publication statusPublished - 2005

Fingerprint

Lycopersicon esculentum
Genetic Recombination
DNA
tomatoes
Chromosomes, Human, Pair 6
Chromosomes, Human, Pair 7
transposition (genetics)
Plant Cells
somatic cells
plasmids
Plasmids
chromosomes
Polymerase Chain Reaction
Genes
genes
cells

Keywords

  • transgenic tomato plants
  • chromosome rearrangements
  • gene cloning
  • recombination
  • genome
  • expression
  • dna
  • identification
  • biotechnology
  • transposition

Cite this

Coppoolse, E. R., de Vroomen, M. J., van Gennip, F., Hersmus, B. J. M., & van Haaren, M. J. (2005). Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites. Plant Molecular Biology, 58(5), 687-698. https://doi.org/10.1007/s11103-005-7705-7
Coppoolse, E.R. ; de Vroomen, M.J. ; van Gennip, F. ; Hersmus, B.J.M. ; van Haaren, M.J. / Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites. In: Plant Molecular Biology. 2005 ; Vol. 58, No. 5. pp. 687-698.
@article{5830d95ebc0d474aabb781af86a568a7,
title = "Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites",
abstract = "Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable.",
keywords = "transgenic tomato plants, chromosome rearrangements, gene cloning, recombination, genome, expression, dna, identification, biotechnology, transposition",
author = "E.R. Coppoolse and {de Vroomen}, M.J. and {van Gennip}, F. and B.J.M. Hersmus and {van Haaren}, M.J.",
year = "2005",
doi = "10.1007/s11103-005-7705-7",
language = "English",
volume = "58",
pages = "687--698",
journal = "Plant Molecular Biology",
issn = "0167-4412",
publisher = "Springer Verlag",
number = "5",

}

Coppoolse, ER, de Vroomen, MJ, van Gennip, F, Hersmus, BJM & van Haaren, MJ 2005, 'Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites', Plant Molecular Biology, vol. 58, no. 5, pp. 687-698. https://doi.org/10.1007/s11103-005-7705-7

Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites. / Coppoolse, E.R.; de Vroomen, M.J.; van Gennip, F.; Hersmus, B.J.M.; van Haaren, M.J.

In: Plant Molecular Biology, Vol. 58, No. 5, 2005, p. 687-698.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Size does matter: Cre-mediated somatic deletion efficiency depends on the distance between the target lox-sites

AU - Coppoolse, E.R.

AU - de Vroomen, M.J.

AU - van Gennip, F.

AU - Hersmus, B.J.M.

AU - van Haaren, M.J.

PY - 2005

Y1 - 2005

N2 - Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable.

AB - Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable.

KW - transgenic tomato plants

KW - chromosome rearrangements

KW - gene cloning

KW - recombination

KW - genome

KW - expression

KW - dna

KW - identification

KW - biotechnology

KW - transposition

U2 - 10.1007/s11103-005-7705-7

DO - 10.1007/s11103-005-7705-7

M3 - Article

VL - 58

SP - 687

EP - 698

JO - Plant Molecular Biology

JF - Plant Molecular Biology

SN - 0167-4412

IS - 5

ER -