초록 ▼

The present disclosure provides for recombinant nucleic acids, and cells and virions comprising the recombinant nucleic acids, that can be used to identify, isolate, and/or purify cells latently infected with immunodeficiency virus. A subject recombinant nucleic acid includes (a) a first nucleotide

The present disclosure provides for recombinant nucleic acids, and cells and virions comprising the recombinant nucleic acids, that can be used to identify, isolate, and/or purify cells latently infected with immunodeficiency virus. A subject recombinant nucleic acid includes (a) a first nucleotide sequence encoding a first reporter polypeptide that produces a first detectable signal, where the first nucleotide sequence is operably linked to an immunodeficiency virus promoter and is translated as an early gene; and (b) a second nucleotide sequence encoding a second reporter polypeptide that produces a second detectable signal that is distinguishable from the first detectable signal, where the second nucleotide sequence is operably linked to a non-immunodeficiency virus promoter. In some aspects, the first and second nucleotide sequences are both positioned between a shared 5′ long terminal repeat (LTR) and a shared 3′ LTR. Also provided are related methods.

대표청구항 ▼

1. A recombinant nucleic acid, comprising: (a) a first nucleotide sequence encoding a first reporter polypeptide that produces a first detectable signal, wherein the first nucleotide sequence is operably linked to an immunodeficiency virus promoter and is translated as an early gene; and(b) a second

1. A recombinant nucleic acid, comprising: (a) a first nucleotide sequence encoding a first reporter polypeptide that produces a first detectable signal, wherein the first nucleotide sequence is operably linked to an immunodeficiency virus promoter and is translated as an early gene; and(b) a second nucleotide sequence encoding a second reporter polypeptide that produces a second detectable signal that is distinguishable from the first detectable signal, wherein the second nucleotide sequence is operably linked to a non-immunodeficiency virus promoter,wherein the first and second nucleotide sequences are both positioned between a shared 5′ long terminal repeat (LTR) and a shared 3′ LTR. 2. The recombinant nucleic acid according to claim 1, wherein the recombinant nucleic acid encodes a transcription-competent immunodeficiency virus and further comprises: a functional transactivation response element (TAR); anda third nucleotide sequence, wherein the third nucleotide sequence encodes a functional transactivator protein (Tat) and is positioned between the shared 5′ long terminal repeat (LTR) and the shared 3′ LTR. 3. The recombinant nucleic acid according to claim 2, wherein the immunodeficiency virus is a human immunodeficiency virus (HIV). 4. The recombinant nucleic acid according to claim 3, wherein the HIV is HIV-1 or HIV-2. 5. The recombinant nucleic acid according to claim 1, wherein the first nucleotide sequence is inserted into the locus of an early gene. 6. The recombinant nucleic acid according to claim 5, wherein the early gene is selected from: nef, tat, and rev. 7. The recombinant nucleic acid according to claim 6, wherein the early gene is nef. 8. The recombinant nucleic acid according to claim 5, wherein the early gene is the early gene that is most proximal to the 3′ LTR. 9. The recombinant nucleic acid according to claim 5, wherein the first nucleotide sequence is inserted in a manner such that the protein encoded by the early gene is not functional or is not produced. 10. The recombinant nucleic acid according to claim 5, wherein the first nucleotide sequence is inserted in a manner such that a protein encoded by the early gene is a fusion protein comprising the protein encoded by the early gene and the first reporter polypeptide. 11. The recombinant nucleic acid of claim 1, wherein at least one of the first and second reporter polypeptides is a fluorescent protein. 12. The recombinant nucleic acid of claim 11, wherein one of the first and second reporter polypeptides is a red fluorescent protein, and the other is a green fluorescent protein. 13. The recombinant nucleic acid of claim 1, wherein the non-immunodeficiency virus promoter is selected from a group consisting of: an EF1α promoter, a Spleen Focus Forming Virus promoter, a ubiquitin promoter, and a cytomegalovirus (CMV) promoter. 14. The recombinant nucleic acid of claim 1, wherein the 5′ LTR comprises the immunodeficiency virus promoter. 15. A virion comprising a recombinant nucleic acid according to claim 1.