Methods and systems for generating nucleic acid diversity
OwnerDavid BIKARD, Raphael LAURENCEAU +1inventors
InventorsDavid BIKARD, Raphael LAURENCEAU, William ROSTAIN
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Abstract
Provided are methods comprising expressing in a recombinant cell a recombinant error-prone reverse transcriptase (RT) and recombinant spacer RNA comprising a target sequence; making a mutagenized cDNA polynucleotide homologous to a DNA sequence in the recombinant cell; expressing a recombinant recombineering system in the recombinant cell; and recombining the mutagenized cDNA with the homologous DNA sequence in the recombinant cell. Also provided are recombinant cells comprising recombinant coding sequences for a recombinant error-prone reverse transcriptase (RT), recombinant spacer RNA comprising a target sequence, and recombinant recombineering system.
Claims20
A method of generating targeted nucleic acid diversity, comprising expressing in a recombinant cell a recombinant error-prone reverse transcriptase (RT) and a recombinant spacer RNA comprising a target sequence;
- making a mutagenized cDNA polynucleotide homologous to a DNA sequence in the recombinant cell;
- expressing a recombinant recombineering system in the recombinant cell;
- and recombining the mutagenized cDNA with the homologous DNA sequence in the recombinant cell.
A recombinant cell comprising recombinant coding sequences for a recombinant error-prone reverse transcriptase (RT) and at least one recombinant spacer RNA comprising a target sequence, and a coding sequence that expresses a recombinant recombineering system.
A kit for generating targeted nucleic acid diversity, comprising one or a plurality of recombinant expression vectors together comprising coding sequences for a recombinant error-prone reverse transcriptase (RT) and at least one recombinant spacer RNA comprising a target sequence, and a coding sequence that expresses a recombinant recombineering system.
Dependencies are inferred from the claim text.