Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-02-22
1998-12-01
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 911, 435 5, 514 44, 514 2, 536 245, 536 243, 536 231, 530300, 530350, C12Q 168, C12P 1934, A61K 4800, C07K 500
Patent
active
058436430
ABSTRACT:
A method and composition are disclosed for transfecting eukaryotic cells using a DNA segment coupled to a site-specific chromosome-binding polypeptide. The polypeptide-DNA conjugate is referred to herein as a polypeptide-linked-rDNA (PLR) molecule. One example of a PLR molecule comprises a DNA segment containing a nucleotide sequence from a normally functioning human gene, coupled by means of a covalent crosslinking reagent to a site-specific chromosome-binding polypeptide (such as a transcription regulating polypeptide that binds to a specific nucleotide sequence in chromosomal DNA). After the PLR molecule enters the cytoplasm of a cell, such as by electroporation, the chromosome-binding polypeptide enables transport of the PLR molecule through the cytoplasm and into the nucleus, using a nuclear localization sequence (NLS) domain of the polypeptide. Inside the nucleus, the polypeptide scans the chromosomes until it binds to a specific chromosomal binding site. This positions the DNA segment of the PLR molecule near a target sequence (such as a defective gene) in the chromosome. The desired DNA segment contained in the PLR molecule has sufficient nucleotide sequence homology with the target gene to enable a recombination event to replace the target gene sequence with the desired gene sequence.
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Jones W. Gary
Kelly Patrick D.
Rees Dianne
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