生物技术通报 ›› 2021, Vol. 37 ›› Issue (9): 266-273.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0015
胡秀文1(), 刘华2,4, 王宇3, 唐雪明1,5(), 王金斌2,4, 曾海娟2,4, 蒋玮2,4, 李红6
收稿日期:
2021-01-04
出版日期:
2021-09-26
发布日期:
2021-10-25
作者简介:
胡秀文,女,硕士研究生,研究方向:分子生物学;E-mail: 基金资助:
HU Xiu-wen1(), LIU Hua2,4, WANG Yu3, TANG Xue-ming1,5(), WANG Jin-bin2,4, ZENG Hai-juan2,4, JIANG Wei2,4, LI Hong6
Received:
2021-01-04
Published:
2021-09-26
Online:
2021-10-25
摘要:
CRISPR-Cas系统是一种高效、实用的基因编辑工具,被广泛应用于基因组编辑和调控机制研究。目前发现的Cas系列核酸酶工具为开发用于各种目的的不同类型核酸检测,具有广阔的应用前景。CRISPR-Cas系统的高灵敏度和高特异性,使其在病原体检测、单核苷酸多态性(SNP)分析以及基因突变检测等发挥了极其重要的作用;另外,CRISPR-Cas系统在核酸检测领域的精确、高效性,间接推动了基础生物学和应用生物学研究的进展。概述了不同类型的靶向特定核酸检测的CRISPR-Cas系统的最新进展和用途,以及与之对应的新一代的体外检测平台,旨为核酸检测领域提供新的研究思路和理论依据。
胡秀文, 刘华, 王宇, 唐雪明, 王金斌, 曾海娟, 蒋玮, 李红. CRISPR-Cas系统在核酸检测中的应用研究[J]. 生物技术通报, 2021, 37(9): 266-273.
HU Xiu-wen, LIU Hua, WANG Yu, TANG Xue-ming, WANG Jin-bin, ZENG Hai-juan, JIANG Wei, LI Hong. Application of CRISPR-Cas System in Nucleic Acid Detection[J]. Biotechnology Bulletin, 2021, 37(9): 266-273.
效应物 Effector | 结构域 Domain | 向导RNA SgRNA | PAM/PFS | 检测平台 Detection platform | 靶向目标 Target type | 信号放大方法 Signal amplification | 灵敏性 Sensitivity | 多路复用 Multiplex | 定量 Quantitative | 便携性 Portable | 时间 Time/h |
---|---|---|---|---|---|---|---|---|---|---|---|
Cas9 | HNH RuvC | tracrRNA | 5'NGG | 无 | DNA | CAS-EXPAR | aM | 无 | 无 | 是 | <1 |
crRNA | |||||||||||
Cas12a Cas12b | RuvC | crRNA | 5'TTN | HOLMES | DNA | PCR; RT-PCR;RPA | aM | 无 | 无 | 否 | 1 |
DETECTR | aM | 无 | 无 | 是 | 1 | ||||||
HOLMESv2 | DNA | LAMP; RT-LAMP | aM | 无 | 有 | 是 | 1 | ||||
Cas13a | HEPN | crRNA | 3'A、U 或C | SHERLOCK | RNA | RPA | aM | 无 | 无 | 是 | 2 |
Cas14 | RuvC | tracrRNA crRNA | 无 | DETECTR | ssDNA | RPA | aM | 无 | 无 | 是 | 1 |
表1 CRISPR检测系统比较
Table 1 Comparison of CRISPR detection system
效应物 Effector | 结构域 Domain | 向导RNA SgRNA | PAM/PFS | 检测平台 Detection platform | 靶向目标 Target type | 信号放大方法 Signal amplification | 灵敏性 Sensitivity | 多路复用 Multiplex | 定量 Quantitative | 便携性 Portable | 时间 Time/h |
---|---|---|---|---|---|---|---|---|---|---|---|
Cas9 | HNH RuvC | tracrRNA | 5'NGG | 无 | DNA | CAS-EXPAR | aM | 无 | 无 | 是 | <1 |
crRNA | |||||||||||
Cas12a Cas12b | RuvC | crRNA | 5'TTN | HOLMES | DNA | PCR; RT-PCR;RPA | aM | 无 | 无 | 否 | 1 |
DETECTR | aM | 无 | 无 | 是 | 1 | ||||||
HOLMESv2 | DNA | LAMP; RT-LAMP | aM | 无 | 有 | 是 | 1 | ||||
Cas13a | HEPN | crRNA | 3'A、U 或C | SHERLOCK | RNA | RPA | aM | 无 | 无 | 是 | 2 |
Cas14 | RuvC | tracrRNA crRNA | 无 | DETECTR | ssDNA | RPA | aM | 无 | 无 | 是 | 1 |
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