Human Cx43 gene inference sequence, shRNA-Cx43 virus and low expression Cx43 protein cell line

Abstract

The invention discloses a human Cx43 gene inference sequence shown as SEQ ID NO1. The invention also discloses a human shRNA-Cx43 virus, which is formed by building the sequence shown as SEQ ID NO1 inthe expression plasmid. The invention also discloses a low expression Cx43 protein cell line, which is a cartilage cell infected by the shRNA-Cx43 virus. The human Cx43 gene interference fragment isbuilt at the downstream part of a U6 promoter of a lentivirus carrier by a molecular biological means. The goals that the targeted gene is interfered, and meanwhile, the green fluorescent protein EGFPand puromycin resistance gene is expressed are achieved; through EGFP, the work state of the carrier can be conveniently observed; through puromycin, the targeted gene interference stable cell straincan be conveniently screened. The built plasmid packaged lentivirus is used for stable strain screening. The stable cell line for low expression connexin43 in the cartilage cell can be used for studying the related inflammatory factor expression quantity change caused by low expression connexin43, so that strategies for effectively treating the arthritis are found.

Description

technical field [0001] The invention relates to human Cx43 gene interference sequence, shRNA-Cx43 virus and cell line with low expression of Cx43 protein. Background technique [0002] Osteoarthritis is a common clinical joint disease and one of the main causes of disability. However, the pathogenesis of osteoarthritis has not been fully elucidated so far. In articular cartilage, connexin43 (Cx43) is the most abundant connexin protein, which plays an important role in maintaining the metabolic homeostasis of articular cartilage, and is considered to be closely related to the occurrence of osteoarthritis. Recent studies have found that the expression of Cx43 is changed in the cartilage damage area of ​​patients with osteoarthritis, and the distribution of Cx43 is changed in the middle and deep non-injury area. junction proteins: differential expression of connexins in normal and osteoarthritic cartilage, 2013). The change of the distribution of Cx43 in the non-injured area...

AI Technical Summary

Problems solved by technology

However, the loss of homotype connexins produces unique pathological phenomena that cannot be fully compensated by other types of connexins (Connexin43 hemichannels mediatesmall molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage, 2014)

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