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Pri-miRNA improved sequence and vector for expressing same

A sequence and carrier technology, applied in the field of RNA interference, can solve the problems of increasing the length of the sequence to be cloned and the difficulty of shRNAmir cloning, and achieve the effect of ensuring high efficiency and accuracy

Pending Publication Date: 2022-04-05
BINZHOU MEDICAL COLLEGE
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Problems solved by technology

This also leads to the fact that each time a clone carries a gene-specific shRNA sequence, a part of the basic sequence of the pri-miRNA is cloned in, which leads to a large increase in the length of the sequence to be cloned, thereby giving shRNA mir The cloning of

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  • Pri-miRNA improved sequence and vector for expressing same
  • Pri-miRNA improved sequence and vector for expressing same
  • Pri-miRNA improved sequence and vector for expressing same

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[0043] It should be pointed out that the following detailed description is exemplary and intended to provide further explanation to the present application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0044] It should be noted that the terminology used here is only for describing specific implementations, and is not intended to limit the exemplary implementations according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form also includes the plural form, and it should also be understood that when the terms "comprising" and / or "comprising" are used in this description, it indicates that there are Features, steps, operations, devices, components and / or combinations thereof.

[0045] The technical solutions of the present invention will be clearly and completely described below...

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Abstract

The invention relates to the technical field of RNA (Ribonucleic Acid) interference, in particular to a pril-miRNA (Ribonucleic Acid) improved sequence based on human miR-30 (Micro Ribonucleic Acid) and a vector for expressing the sequence, and BamHI and Apal restriction endonuclease recognition sites are introduced into a stem part of the pril-miRNA improved sequence. According to the invention, a sequence of pril-miRNA, namely a frame of shRNAmir, is modified, two ends of a gene-specific shRNA sequence of a stem part of the pril-miRNA are improved on the premise of not influencing biological processing of the pril-miRNA, and a part of the original sequence is replaced by BamHI and ApaI restriction enzyme recognition sites, namely, only the shRNA sequence is arranged between the recognition sites of the two restriction enzymes, namely BamHI and ApaI, so that only the shRNA sequence is arranged between the recognition sites of the two restriction enzymes; therefore, it is guaranteed that only a very short gene specific shRNA sequence is cloned in the cloning process, the latter can be synthesized by using a DNA oligonucleotide chemical synthesis technology, the problem that a pril-miRNA basic sequence is cloned in is effectively avoided, and high efficiency and accuracy of cloning are guaranteed.

Description

technical field [0001] The invention relates to the technical field of RNA interference, in particular to an improved human miR-30-based pri-miRNA sequence and a vector expressing the sequence. Background technique [0002] RNA interference technology refers to the phenomenon of highly conserved, highly efficient and specific degradation of homologous mRNA induced by double-stranded RNA in the evolution process. It is currently a widely used cell biology technology to reduce the expression level of target genes. [0003] The early RNA interference technology directly interferes with the translation of endogenous mRNA by introducing exogenous siRNA into eukaryotic cells, thereby achieving the effect of RNA interference. However, due to the high cost of this method and the inability to stably interfere with the target gene, researchers have developed the shRNA technology. shRNA technology refers to the expression of shRNA with a hairpin structure through a DNA vector, which g...

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Application Information

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IPC IPC(8): C12N15/113C12N15/65C12N15/867
Inventor 王大鹏
Owner BINZHOU MEDICAL COLLEGE
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