Self-diversifying systems and methods of use thereof
A diversity-generating system comprising: a polynucleotide comprising: a variable region within a sequence encoding a protein or polypeptide, wherein the system is configured to generate a library of diversified molecules with an efficiency of 80% as measured by number of the diversified molecules in total number of molecules. The system may further comprise a template region; and a reverse transcriptase or functional domain thereof.
1 . A self-diversifying system comprising an in vitro engineered vector, wherein the vector comprises a promoter, a polynucleotide sequence, and a nucleic acid template region, wherein the polynucleotide sequence is operably linked to the promoter, is the nucleotide sequence selected from SEQ ID Nos. 98-123, and comprises one or more open reading frames, wherein at least one of the open reading frames comprises a self-diversifying element, wherein the nucleic acid template region is operably linked to a nucleic acid variable region of the polynucleotide sequence, and wherein the self-diversifying element is a diversity generating retroelement (DGR) or a non-DGR.
2 . The system of claim 1 , wherein the self-diversifying element is a (DGR).
3 . The system of claim 1 , wherein the self-diversifying element is a non-DGR.
4 . The system of claim 1 , further comprising a reverse transcriptase or functional domain thereof.
5 . The system of claim 1 , wherein the polynucleotide sequence encodes c-type lectin fold or a fragment thereof.
6 . The system of claim 1 , wherein the polynucleotide sequence encodes a protein or polypeptide capable of binding to a protein or a nucleic acid.
7 . The system of claim 1 , wherein the nucleic acid variable region and the nucleic acid template region have a sequence homology less than 80%.
8 . The system of claim 1 , wherein the system is capable of generating a population of proteins.
9 . The system of claim 1 , wherein the system is capable of generating a population of polynucleotides.
10 . The system of claim 1 , wherein the system is capable of generating one or more binding partners of a target molecule.
11 . The system of claim 10 , wherein the target molecule is a protein.
12 . The system of claim 10 , wherein the target molecule is an antigen.
13 . The system of claim 10 , wherein the target molecule is a polynucleotide.
14 . The system of claim 1 , wherein the system generates a library of diversified molecules when the vector is introduced to cells.
15 . The system of claim 14 , wherein the diversified molecules are polynucleotide-binding proteins or polypeptide-binding proteins.
16 . The system of claim 15 , wherein the polynucleotide-binding proteins or polypeptide-binding proteins are selected from SEQ ID NOs: 3 and 4.
17 . The system of claim 16 , wherein SEQ ID NOs: 3 and 4 are encoded by SEQ ID NO: 1 and 2 respectively.
18 . The system of claim 1 , wherein the nucleic acid variable region is within an open reading frame of the one or more open reading frames.
19 . The system of claim 1 , wherein the self-diversifying element comprises one or more initiation of mutagenic homing (IMH) sequences.