基因组精准编辑技术及其在林木育种中的应用

doi:10.12302/j.issn.1000-2006.202402006

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南京林业大学学报（自然科学版） ›› 2025, Vol. 49 ›› Issue (1): 11-20.doi: 10.12302/j.issn.1000-2006.202402006

• 林学前沿（执行主编尹佟明陈金慧） • 上一篇下一篇

基因组精准编辑技术及其在林木育种中的应用

姜波(), 安新民^*()

林木遗传育种全国重点实验室,林木育种与生态修复国家工程中心,林木分子设计育种高精尖创新中心,林木花卉遗传育种教育部重点实验室,树木花卉育种生物工程国家林业和草原局重点实验室,北京林业大学生物科学与技术学院,北京 100083

收稿日期:2024-02-07 修回日期:2024-10-15 出版日期:2025-01-30 发布日期:2025-01-21
通讯作者: * 安新民(anxinmin@bjfu.edu.cn),教授。
作者简介:
姜波(961788757@qq.com),博士生。
基金资助:
国家重点研发计划(2021YFD2200101-03);科技创新2030-重大项目(2022ZD0401503);国家自然科学基金项目(31870652)

Precise genomic editing technology and its application in the improvement of woody plants

JIANG Bo(), AN Xinmin^*()

State Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, Advanced Innovation Center for Tree Breeding by Molecular Design, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education, Tree and Ornamental Plant Breeding and Biotechnology Laboratory of National Forestry and Grassland Administration, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China

Received:2024-02-07 Revised:2024-10-15 Online:2025-01-30 Published:2025-01-21

摘要/Abstract

摘要：

基因组精准编辑技术正在重新定义对生命奥秘的理解,其核心在于能够在特定的基因组位点精确地插入、删除或替换DNA序列,从而实现对生物体内遗传信息的定向精准编辑。这些技术已经成为现代生物领域研究的基石,从早期的探索到CRISPR系统的开创性应用,每一步都展示了科学探索的深远影响。CRISPR系统的应用带来了基因组编辑的巨大飞跃,它催化了更精准的编辑工具出现,如碱基编辑器和先导编辑器等,它们显著增强了精准编辑基因组的能力。这一历史性转变已经在农业改良、疾病治疗等领域展现出巨大的潜力。基因编辑技术的应用前景广阔,CRISPR/Cas系统能敲除多个基因,具有高靶向效率、易于设计和操作,且成本较低等优点,因此在作物、林木遗传改良中被广泛应用;但它也有不少不足之处,而每一款新编辑工具的出现都使这一技术得以不断完善。随着技术的不断进步,基因编辑技术有望在未来解决更多复杂的生物学问题,为人类健康和农林业发展带来更多的创新和突破力。

关键词: CRISPR, Cas9, 胞嘧啶碱基编辑器, 腺嘌呤碱基编辑器, 糖基化酶碱基编辑器, 双碱基编辑器, 先导编辑器

Abstract:

The rapid advancement of genome precision editing technologies has revolutionized our understanding of life’s mysteries. Central to these technologies is the capacity to precisely insert, delete, and substitute DNA sequences at designated genomic positions, facilitating the targeted modification of genetic information within living organisms. These technologies have become the cornerstone of research in the field of biology, from early explorations to the groundbreaking application of the CRISPR system, emphasizing the profound impact of scientific inquiry. The application of the CRISPR system has brought about a significant leap in genomic editing, catalyzing the emergence of more precise editing tools, such as base editors and prime editors, which have significantly enhanced the ability to edit genomes with precision. This milestone shift has already demonstrated tremendous potential in a wide range of fields, including agricultural improvement and disease treatment. The potential applications for gene editing technology are vast, especially in forestry and grass breeding. The CRISPR/Cas system can knock out multiple genes, offering high targeting efficiency, ease of design and operation, and low costs, making it widely applicable in crop genetic improvement. However, it is also associated with numerous limitations, which can be overcomed by new editing tools. As the technologies continues to advance, gene editing technology is expected to solve complex biological problems in the future, bringing more innovation and breakthroughs to human health and agricultural development.

Key words: CRISPR, Cas9, cytosine base editor, adenine base editor, glycosylase base editor, dual base editor, prime editor

中图分类号:

S722.3

姜波,安新民. 基因组精准编辑技术及其在林木育种中的应用[J]. 南京林业大学学报（自然科学版）, 2025, 49(1): 11-20.

JIANG Bo, AN Xinmin. Precise genomic editing technology and its application in the improvement of woody plants[J].Journal of Nanjing Forestry University (Natural Science Edition), 2025, 49(1): 11-20.DOI: 10.12302/j.issn.1000-2006.202402006.

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表3

精准编辑技术在疾病治疗与植物育种等方面的一些应用"

编辑器类型 editor type	优点 advantage		不足 limitation		物种 species		应用 application		参考文献 reference
胞嘧啶碱基编辑器 CBE	编辑效率相对较高 high editing efficiency		只能进行几种碱基转换 only a few base transitions can be performed 脱靶效应高 high off target effect 编辑范围局限 limitations of editing scope		小鼠 Mus musculus		胆固醇疾病 cholesterol disorders		[36]
					猕猴 Macaca mulatta		胆固醇疾病 cholesterol disorders		[37]
					人类 Homo sapiens		β-地中海贫血症 β-thalassemia		[39]
							马凡综合征 marfan syndrome		[40]
					水稻 Oryza sativa		ALS基因		[41]
					玉米 Zea mays		ALS基因		[42]
					小麦 Triticum aestivum		ALS基因		[43]
							ACCase基因		[44]
					烟草 Nicotiana tabacum		ALS基因		[45]
					甜橙 Citrus sinensis		ALS基因		[46]
					杨树 Populus		Platz基因		[47]
腺嘌呤碱基编辑器 ABE					小鼠 Mus musculus		Ⅰ型酪氨酸血症 yype Ⅰ tyrosinemia		[48]
							杜氏肌营养不良症 duchenne muscular dystrophy		[38]
							Ⅰ型黏多糖病 type Ⅰ mucopolysaccharidosis		[49]
							早衰症 progeria		[50]
					水稻 O. sativa		ACCase基因		[51]
					甜橙 C. sinensis		溃疡病 ulcer		[46]
					葡萄柚 C. paradise		溃疡病 ulcer		[46]
糖基化酶碱基编辑器GBE					水稻 O. sativa		ALS基因		[52]
先导编辑器 PE		能够实现12种碱基替换 12 types of base substitutions can be achieved 多碱基替换 multiple base substitutions 小片段的插入或删除 insertion or deletion of small fragments 脱靶效应低 low off target effect 编辑范围广 wide editing scope		编辑效率相对较低 low editing efficiency 使用设计过程复杂 complex design process		人类 H. sapiens		杜氏肌营养不良症 duchenne muscular dystrophy		[38]
						小鼠 Mus musculus		Ⅰ型酪氨酸血症 yype Ⅰ tyrosinemia		[48]
								雷柏氏先天性黑矇症 leber’s congenital melanosis		[53]
						水稻 O. sativa		ACCase基因		[54]
						玉米 Z. mays		ALS基因		[55]
								ACCase基因		[55]

表3

参考文献 62

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胞嘧啶碱基编辑器 cytosine base editor	PAM序列 PAM sequence	编辑器结构 editor structure	NLS个数 number of NLS	活性窗口位置 active window position	参考文献 reference
BE1	NGG	rAPOBEC1-dCas9	1	4—8	[14]
BE2	NGG	rAPOBEC1-dCas9-UGI	1	4—8	[14]
BE3	NGG	rAPOBEC1-nCas9(D10A) -UGI	1	4—8	[14]
hA3A-BE3	NGG	hAPOBEC3A-nCas9(D10A)-UGI	1	2—13	[17]
BE3-NG	NG	rAPOBEC1-nCas9(D10A)-NG-UGI	1	4—8	[21]
Target-AID	NGG	nCas9(D10A)-PmCDA1-UGI	1	2—8	[18]
BE4	NGG	rAPOBEC1-nCas9(D10A)-UGI-UGI	1	4—8	[22]
BE4max	NGG	rAPOBEC1-nCas9(D10A)-UGI-UGI	2	4—8	[20]
TAM	NGG	dCas9-hAIDx	1	4—10	[19]
xBE3	NGN、GAA、GAT	rAPOBEC1-nxCas9(D10A)-UGI	1	4—8	[23]

腺嘌呤碱基编辑器 adenine base editor	PAM序列 PAM sequence	编辑器结构 editor structure	NLS个数 number of NLS	活性窗口位置 active window position	参考文献 reference
ABE7.10	NGG	TadA-TadA^*-nCas9(D10A)	1	4—7	[24]
xABE	NGN、GAA、GAT	TadA-TadA^*-nxCas9(D10A)	1	4—7	[23]
xABEmax	NGN、GAA、GAT	TadA-TadA^*-nxCas9(D10A)	2	4—8	[25]

基因组精准编辑技术及其在林木育种中的应用

Precise genomic editing technology and its application in the improvement of woody plants

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