CRISPR/Lbcpf1 gene editing system and application thereof

A gene editing and gene silencing technology, applied in the fields of bioengineering and biomedicine, which can solve problems such as toxic side effects, treatment obstacles, and unbearable patients.

Pending Publication Date: 2021-06-11
鼓润(武汉)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, chemotherapy and radiotherapy have certain complications, mainly radiation-induced rectum and cystitis, which can only be recovered slowly after active treatment and

Method used

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  • CRISPR/Lbcpf1 gene editing system and application thereof
  • CRISPR/Lbcpf1 gene editing system and application thereof
  • CRISPR/Lbcpf1 gene editing system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Immunohistochemical technique detects the expression of HMGA2 protein in different tissue samples

[0033] In this embodiment, the expression of HMGA2 protein in different tissue samples was detected by histology chip.

[0034] Select the tissue samples of different degrees of cervical lesions in the histological chips CIN481 and CIN1201 listed in Table 1 (the histological chips CR501 and BC10025 are used as controls) (according to the degree of lesions from mild to severe, they are divided into: normal or inflammatory, CIN I, CIN II, CIN III, SCC, ACC) paraffin sections were detected by immunohistochemistry to detect the expression of HMGA2 protein.

[0035] Table 1: Histological information in histology chips CIN481, CIN1201, CR501 and BC10025

[0036]

[0037]

[0038] *Severity of Cervical Intraepithelial Neoplasia (CIN)

[0039] Select paraffin sections of tissue samples with different degrees of cervical lesions in the above-mentioned histolo...

Embodiment 2

[0052] Embodiment 2: Immunocytochemical technique detects the expression of HMGA2 protein in different cell samples

[0053] The cell samples used in this example were from HPV-positive patients undergoing cervical cancer screening in the First Affiliated Hospital of Sun Yat-sen University from November 2016 to December 2017. Such as image 3 As shown, the obtaining process of the cell sample adopted in this embodiment is shown: the patient accepts HPV (detecting whether to be infected with human papillomavirus) and LCT (cervical lesion cytology detection) detection at the same time, or they have a sample from another authoritative medical institution HPV test results. Exclusion criteria were women over 65 years of age, repeatedly tested or treated. 227 HPV-positive women were enrolled (approved by the Ethics Committee of the First Affiliated Hospital of Sun Yat-Sen University (No. [2016] 177)) and treated according to the 2016 American College of Obstetrics and Gynecology (...

Embodiment 3

[0068] Embodiment 3: Comparison of HMGA2 and LCT detection efficiency

[0069] According to the method of Example 2, colposcopy was performed on 227 HPV-positive patients who were screened for cervical cancer in the First Affiliated Hospital of Sun Yat-sen University from November 2016 to December 2017 (obtained from the First Affiliated Hospital of Sun Yat-sen University). Approval of the Ethics Committee (No.[2016]177)), finally obtained 77 cases of HPV detection and colposcopy fully matched cell samples, as this example using molecular biology methods (HMGA2) and using cytology detection methods (LCT ) cervical cell samples to evaluate the efficiency of HMGA2 protein or LCT as a single index to detect CIN II+ in HPV positive patients. The above 77 cell samples were made into cell smears and stored in BD SurePath LBC preservative solution (BD Diagnostics, Sparks, MD) for future use, and HMGA2 index detection and LCT index detection were respectively carried out according to ...

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Abstract

The invention discloses a CRISPR/Lbcpf1 gene editing system and an application thereof, belongs to the field of bioengineering and biomedicine, and relates to early prevention and treatment of cervical lesions. The CRISPR/Lbcpf1 gene editing system provided by the invention comprises crRNA expression plasmids and Lbcpf1 expression plasmids, the crRNA expression plasmids and the Lbcpf1 expression plasmids are co-transfected into cells to express and form an Lbcpf1-crRNA binary complex, and the recognition and shearing of an HMGA2 gene can be realized, so that the function of the gene is lost; the CRISPR/Lbcpf1 gene editing system can specifically cause apoptosis of HPV positive cervical lesion cells, and can be used as a gene therapy drug for effectively treating HPV positive cervical lesion.

Description

technical field [0001] The invention belongs to the fields of bioengineering and biomedicine, and relates to the early prevention and treatment of cervical lesions, in particular to a CRISPR / Lbcpf1 gene editing system and its application, especially in silencing HMGA2 gene and preparing medicines for treating HPV-positive cervical lesions Applications. Background technique [0002] Cervical cancer is one of the most common gynecological malignancies in women all over the world after breast cancer. Its incidence rate ranks second among female malignancies, and its incidence rate ranks first in some developing countries. According to the International Cancer Research Center, there are about 527,600 new cases of cervical cancer each year, accounting for about 7.9% of all new cancer cases in women, 84% of which occur in developing countries; there are about 265,700 cervical cancer deaths each year , accounting for approximately 7.5% of all new cancer cases in women, 87% of whic...

Claims

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Application Information

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IPC IPC(8): C12N15/90C12N15/85A61K31/7105A61K48/00A61P35/00
CPCC12N15/907C12N15/85C07K14/47A61K31/7105A61P35/00C12N2310/20C12N2800/107
Inventor 谢红娴
Owner 鼓润(武汉)医疗科技有限公司
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