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1. WO2020118169 - MATRIX ATTACHMENT REGIONS AND USES IN PROMOTING GENE EXPRESSION

Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

[ EN ]

WHAT IS CLAIMED IS:

1. A recombinant polynucleotide comprising a coding sequence, a promoter configured to initiate the transcription of the coding sequence, and a matrix attachment region (MAR) core selected from the group consisting of SEQ ID NO: 1, 5, 9 and 13, and nucleic acid sequences having at least 90% sequence identity to any one of SEQ ID NO: 1, 5, 9 and 13, wherein the MAR core is capable to attach to a mammalian nuclear matrix.

2. The polynucleotide of claim 1, further comprising a 5’ flanking region selected from SEQ ID NO: 2, 6, 10, and 14 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 2, 6, 10, and 14.

3. The polynucleotide of claim 2, wherein the 5’ flanking region is 5’ to, and within 100 nucleotides from, the MAR core.

4. The polynucleotide of any one of claims 1-3, further comprising a 3’ flanking region selected from SEQ ID NO: 3, 7, 11, and 15 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 3, 7, 11, and 15.

5. The polynucleotide of claim 4, wherein the 3’ flanking region is 3’ to, and within 100 nucleotides from, the MAR core.

6. The polynucleotide of claim 1, which comprises a MAR selected from SEQ ID NO: 4, 8, 12 and 16, and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 4, 8, 12 and 16, wherein the MAR is capable to attach to a mammalian nuclear matrix.

7. A cell comprising the polynucleotide of any one of claims 1-6.

8. A method of transfecting to a cell a coding sequence, comprising contacting the cell with a first polynucleotide comprising the coding sequence and a promoter for initiating transcription of the coding sequence, and a second, unlinked polynucleotide comprising a matrix attachment region (MAR) core selected from SEQ ID NO: 1, 5, 9 and 13, and nucleic acid sequences having at least 90% sequence identity to any one of SEQ ID NO: 1, 5, 9 and 13, wherein the MAR core is capable to attach to a mammalian nuclear matrix, under conditions for the first and second polynucleotides to transfect into the cell.

9. The method of claim 8, wherein the second polynucleotide further comprises a 5’ flanking region selected from SEQ ID NO: 2, 6, 10, and 14 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 2, 6, 10, and 14.

10. The method of claim 9, wherein the 5’ flanking region is 5’ to, and within 100 nucleotides from, the MAR core.

11. The method of any one of claims 8-10, wherein the second polynucleotide further comprises a 3’ flanking region selected from SEQ ID NO: 3, 7, 11, and 15 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 3, 7, 11, and 15 .

12. The method of claim 11, wherein the 3’ flanking region is 3’ to, and within 100 nucleotides from, the MAR core.

13. The method of claim 8, wherein the second polynucleotide further comprises a MAR selected from SEQ ID NO: 4, 8, 12 and 16, and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 4, 8, 12 and 16, wherein the MAR is capable to attach to a mammalian nuclear matrix.

14. A transfected cell prepared by the method of any one of claims 8-13.

15. A chimeric matrix attachment region (MAR), comprising:

(a) a MAR core selected from SEQ ID NO: 1, 5, 9 and 13, and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 1, 5, 9 and 13, wherein the MAR core is capable to attach to a mammalian nuclear matrix,

(b) a 5’ flanking region selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30 and 34 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 2, 6, 10,

14, 18, 22, 26, 30 and 34, and

(c) a 3’ flanking region selected from SEQ ID NO: 3, 7, 11, 15, 19, 23, 27, 31 and 35 and nucleic acid sequences having at least 75% sequence identity to any one of SEQ ID NO: 3, 7, 11,

15, 19, 23, 27, 31 and 35,

wherein the MAR core, the 5’ flanking region and the 3’ flanking region are not from the same natural MAR.