Vector system for expressing regulatory RNA
Viral vectors, lentiviral particles, and modified cells are disclosed. They encode or express a small RNA capable of targeting the KIF11 gene. In embodiments, the viral vectors and lenti viral particles further comprise and a KIF11 gene whose non-coding region has been modified such that it is resistant to activity by the small RNA.
1. A modified mesenchymal stem cell comprising a mesenchymal stem cell infected with a lentiviral particle, wherein the lentiviral particle comprises:
an envelope protein capable of infecting the mesenchymal stem cell; and
a first nucleotide sequence encoding a small RNA capable of binding a non-coding region of a host copy of KIF11,
wherein the small RNA comprises a sequence having at least 80% sequence identity with at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
2. The modified mesenchymal stem cell of claim 1 , wherein the small RNA comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
3. A method of producing a modified mesenchymal stem cell, the method comprising:
infecting a mesenchymal stem cell with an effective amount of a lentiviral particle, wherein the lentiviral particle comprises:
an envelope protein capable of infecting the mesenchymal stem cell; and
a first nucleotide sequence encoding a small RNA capable of binding a non-coding region of a host copy of KIF11,
wherein the small RNA comprises a sequence having at least 80% sequence identity with at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
4. The method of claim 3 , wherein the small RNA comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
5. A viral vector comprising a first nucleotide sequence encoding a small RNA capable of binding a non-coding region of a host copy of KIF11,
wherein the small RNA comprises a sequence having at least 80% sequence identity with at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
6. The viral vector of claim 5 , wherein the small RNA comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
7. A lentiviral particle produced by a packaging cell and capable of infecting a target cell, the lentiviral particle comprising:
an envelope protein capable of infecting the target cell; and
a first nucleotide sequence encoding a small RNA capable of binding a non-coding region of a host copy of KIF11,
wherein the small RNA comprises a sequence having at least 80% sequence identity with at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.
8. The lentiviral particle of claim 7 , wherein the small RNA comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.