Templated mutagenesis and nucleic acid synthesis
AssigneeThe Trustees of Columbia University in the City of New York
InventorsHarris Wang, Liyuan Liu
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Abstract
The present disclosure provides to systems, methods, and compositions for template-based DNA, synthesis. Particularly the present disclosure provides systems, methods, and compositions for generating multi-site (e.g., three or more) sequence variants of template nucleic acid strands kilobases (e.g., greater than Ikb) m length.
Claims32
A method for modifying a nucleic acid comprising:
- annealing three or more mutagenic primers to a template nucleic acid, wherein each of the three or more mutagenic primers comprises at least one mutagenic nucleotide in comparison to template strand;
- and
- contacting primed template nucleic acid with a polymerase, a ligase, and nucleotide triphosphates under conditions suitable to extend and ligate a first mutant strand.
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A polypeptide comprising a viral capsid protein having two or more amino acid insertions between positions selected from:
- 37/38, 139/140, 190/191, 447/448, 501/502, and 591/592 relative to SEQ ID NO:
- 509,
- wherein each of the two or more amino acid insertions is individually a negatively charged amino acid.
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Dependencies are inferred from the claim text.