Mechanism of chromosomal integration of transgenes in microinjected mouse eggs: sequence analysis of genome-transgene and transgene-transgene junctions at two loci

Tsuyoshi Hamada, Hiroyuki Sasaki, Ritsuko Seki, Yoshiyuki Sakaki

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Production of transgenic animals is a key technique in modern biology, but the process of chromosomal integration of transgenes in microinjected eggs is still not fully understood. To gain information on the mechanisms involved in this process, we have cloned two transgene loci and their corresponding pre-integration sites and compared the junction sequences with the parental nucleotide (nt) sequences. No extensive DNA rearrangements were detected at these loci: only simple deletions (caused by the integration of the transgene concatemers) were present in the host genome. Analysis of three transgene-transgene junctions within the concatemers showed that 'nibbling' of ends (up to 3 nt) had occurred at some ends prior to joining. At all four genome-transgene junctions, short homologies of 1 to 3 nt were found, and at least three of these junctions were associated with the consensus sequence for topoisomerase-I cleavage sites. Moreover, three of the four integration junctions occurred in the terminal regions of the injected sequence, at positions only a few nt away from the ends. These results suggest that linear, but not circular, concatemers were preferentially integrated at their ends utilizing short homologies to the host genome.

Original languageEnglish
Pages (from-to)197-202
Number of pages6
JournalGene
Volume128
Issue number2
DOIs
Publication statusPublished - Jun 30 1993

Fingerprint

Transgenes
Eggs
Sequence Analysis
Genome
Nucleotides
Type I DNA Topoisomerase
Genetically Modified Animals
Gene Rearrangement
Consensus Sequence

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Mechanism of chromosomal integration of transgenes in microinjected mouse eggs : sequence analysis of genome-transgene and transgene-transgene junctions at two loci. / Hamada, Tsuyoshi; Sasaki, Hiroyuki; Seki, Ritsuko; Sakaki, Yoshiyuki.

In: Gene, Vol. 128, No. 2, 30.06.1993, p. 197-202.

Research output: Contribution to journalArticle

@article{0edefb39c6df4e9d8d3a05f317eb8c4a,
title = "Mechanism of chromosomal integration of transgenes in microinjected mouse eggs: sequence analysis of genome-transgene and transgene-transgene junctions at two loci",
abstract = "Production of transgenic animals is a key technique in modern biology, but the process of chromosomal integration of transgenes in microinjected eggs is still not fully understood. To gain information on the mechanisms involved in this process, we have cloned two transgene loci and their corresponding pre-integration sites and compared the junction sequences with the parental nucleotide (nt) sequences. No extensive DNA rearrangements were detected at these loci: only simple deletions (caused by the integration of the transgene concatemers) were present in the host genome. Analysis of three transgene-transgene junctions within the concatemers showed that 'nibbling' of ends (up to 3 nt) had occurred at some ends prior to joining. At all four genome-transgene junctions, short homologies of 1 to 3 nt were found, and at least three of these junctions were associated with the consensus sequence for topoisomerase-I cleavage sites. Moreover, three of the four integration junctions occurred in the terminal regions of the injected sequence, at positions only a few nt away from the ends. These results suggest that linear, but not circular, concatemers were preferentially integrated at their ends utilizing short homologies to the host genome.",
author = "Tsuyoshi Hamada and Hiroyuki Sasaki and Ritsuko Seki and Yoshiyuki Sakaki",
year = "1993",
month = "6",
day = "30",
doi = "10.1016/0378-1119(93)90563-I",
language = "English",
volume = "128",
pages = "197--202",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Mechanism of chromosomal integration of transgenes in microinjected mouse eggs

T2 - sequence analysis of genome-transgene and transgene-transgene junctions at two loci

AU - Hamada, Tsuyoshi

AU - Sasaki, Hiroyuki

AU - Seki, Ritsuko

AU - Sakaki, Yoshiyuki

PY - 1993/6/30

Y1 - 1993/6/30

N2 - Production of transgenic animals is a key technique in modern biology, but the process of chromosomal integration of transgenes in microinjected eggs is still not fully understood. To gain information on the mechanisms involved in this process, we have cloned two transgene loci and their corresponding pre-integration sites and compared the junction sequences with the parental nucleotide (nt) sequences. No extensive DNA rearrangements were detected at these loci: only simple deletions (caused by the integration of the transgene concatemers) were present in the host genome. Analysis of three transgene-transgene junctions within the concatemers showed that 'nibbling' of ends (up to 3 nt) had occurred at some ends prior to joining. At all four genome-transgene junctions, short homologies of 1 to 3 nt were found, and at least three of these junctions were associated with the consensus sequence for topoisomerase-I cleavage sites. Moreover, three of the four integration junctions occurred in the terminal regions of the injected sequence, at positions only a few nt away from the ends. These results suggest that linear, but not circular, concatemers were preferentially integrated at their ends utilizing short homologies to the host genome.

AB - Production of transgenic animals is a key technique in modern biology, but the process of chromosomal integration of transgenes in microinjected eggs is still not fully understood. To gain information on the mechanisms involved in this process, we have cloned two transgene loci and their corresponding pre-integration sites and compared the junction sequences with the parental nucleotide (nt) sequences. No extensive DNA rearrangements were detected at these loci: only simple deletions (caused by the integration of the transgene concatemers) were present in the host genome. Analysis of three transgene-transgene junctions within the concatemers showed that 'nibbling' of ends (up to 3 nt) had occurred at some ends prior to joining. At all four genome-transgene junctions, short homologies of 1 to 3 nt were found, and at least three of these junctions were associated with the consensus sequence for topoisomerase-I cleavage sites. Moreover, three of the four integration junctions occurred in the terminal regions of the injected sequence, at positions only a few nt away from the ends. These results suggest that linear, but not circular, concatemers were preferentially integrated at their ends utilizing short homologies to the host genome.

UR - http://www.scopus.com/inward/record.url?scp=0027256614&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027256614&partnerID=8YFLogxK

U2 - 10.1016/0378-1119(93)90563-I

DO - 10.1016/0378-1119(93)90563-I

M3 - Article

C2 - 8390388

AN - SCOPUS:0027256614

VL - 128

SP - 197

EP - 202

JO - Gene

JF - Gene

SN - 0378-1119

IS - 2

ER -