Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites

被引：13

作者：

Stewart, S ^{[1
]}

MacDonald, N ^{[1
]}

Perkins, E ^{[1
]}

deJong, G ^{[1
]}

Perez, C ^{[1
]}

Lindenbaum, M ^{[1
]}

机构：

[1] Chromos Mol Syst Inc, Burnaby, BC V5A 1W9, Canada

来源：

GENE THERAPY | 2002年 / 9卷 / 11期

关键词：

mammalian artificial chromosomes; ACes; loxP; RFP; gene therapy;

D O I：

10.1038/sj.gt.3301757

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A satellite DNA-based mammalian artificial chromosome (ACes) was generated and subsequently modified by targeting of a loxP-red fluorescent protein (RFP) expression cassette via homologous recombination into a ribosomal DNA (rDNA)-containing locus. Clones containing correctly targeted ACes were identified by PCR from populations of RFP-expressing cells enriched by FACS sorting and were further characterized by fluorescent in situ hybridization. The targeted ACes maintained its ability to be purified to near homogeneity. Studies are currently underway to further characterize the functionality, carrying capacity, stability and transfectability of this modified ACes.

引用

页码：719 / 723

页数：5

共 19 条

[1] DOES T-LOCUS IN MOUSE INCLUDE RIBOSOMAL DNA [J].

ATWOOD, KC ;

GLUECKSOHNWAELSCH, S ;

YU, MT ;

HENDERSON, AS .

CYTOGENETICS AND CELL GENETICS, 1976, 17 (01) :9-17

[2]

CHEN CA, 1988, BIOTECHNIQUES, V6, P632

[3] Generation of transgenic mice and germline transmission of a mammalian artificial chromosome introduced into embryos by pronuclear microinjection [J].

Co, DO ;

Borowski, AH ;

Leung, JD ;

van der Kaa, J ;

Hengst, S ;

Platenburg, GJ ;

Pieper, FR ;

Perez, CF ;

Jirik, FR ;

Drayer, JI .

CHROMOSOME RESEARCH, 2000, 8 (03) :183-191

[4]

Csonka E, 2000, J CELL SCI, V113, P3207

[5] Efficient in-vitro transfer of a 60-Mb mammalian artificial chromosome into murine and hamster cells using cationic lipids and dendrimers [J].

de Jong, G ;

Telenius, A ;

Vanderbyl, S ;

Meitz, A ;

Drayer, J .

CHROMOSOME RESEARCH, 2001, 9 (06) :475-485

[6]

deJong G, 1999, CYTOMETRY, V35, P129, DOI 10.1002/(SICI)1097-0320(19990201)35:2<129::AID-CYTO4>3.0.CO

[7]

2-A

[8] Repeat-induced gene silencing in mammals [J].

Garrick, D ;

Fiering, S ;

Martin, DIK ;

Whitelaw, E .

NATURE GENETICS, 1998, 18 (01) :56-59

[9] Stable expression of green fluorescent protein after liposomal transfection of K562 cells without selective growth conditions [J].

Gubin, AN ;

Koduru, S ;

Njoroge, JM ;

Bhatnagar, R ;

Miller, JL .

BIOTECHNIQUES, 1999, 27 (06) :1162-+

[10]

Hadlaczky G, 2001, CURR OPIN MOL THER, V3, P125

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