引导编辑系统研究进展.pdfVIP

引导编辑系统研究进展

摘要:引导编辑(Prime

editing，PE)系统是一种全新的、革命性的基因组编辑策略。该系统由引导编辑器(Prime

editor)组成，包括nCas9(H840A)与逆转录酶(Reverse

transcriptase，RT)的融合蛋白；以及包含PBS(Primer

binding

site)序列和RT模板(RT

template，RTT)序列的pegRNA(Prime

editing

guide

RNA)两大部分。PE系统

可以在双链不断裂的情况下实现所有12种类型的碱基替换及小片段DNA增删，是精准编辑的全新范式。自

2019年开发至今不到4年时间，PE系统作为一种通用的技术平台，已广泛应用于医疗、农业等各个领域，产生

了一大批新种质资源、基因治疗药物等优秀应用案例。PE作为目前最灵活、最具发展前景的基因组精准编辑新

手段，仍旧存在效率偏低、大片段操纵能力不足、系统组分设计复杂(如pegRNA)、安全性未全面评估等问题，仍

需要深入研究。本文详细介绍了PE系统的技术原理及限制因素，全面总结了PE系统自开发以来的优化策略及

在动植物系统、医疗领域的应用现状，并对PE的发展前景进行了展望。

关键词:引导编辑系统；优化策略；农业应用；医疗应用

Recentadvancesinprimeeditingsystem

Abstract:Prime

editing

(PE)

system

is

a

newly

developed

and

greatly

revolutionized

genome

editing

technology.

The

system

is

based

on

prime

editors,

which

are

composed

of

two

components:

A

fusion

protein

of

nCas9

(H840A)

and

reverse

transcriptase

(RT),

and

a

pegRNA

which

contains

a

PBS

(Primer

binding

site)

sequence

and

an

RT

template

(RTT)

sequence.

The

PE

system

can

realize

all

12

types

of

base

substitutions

and

small

fragment

DNA

additions

and

deletions

without

double-strand

breaks,

which

is

a

new

paradigm

for

precision

editing.

In

less

than

4

years

since

its

development

in

2019,

the

PE

system,

as

a

universal

technology

platform,

has

been

widely

used

in

various

fields

such

as

healthcare

and

agriculture,

generating

a

large

number

of

excellent

application

cases

such

as

new

germplasm

resources

and

gene

therapy

drugs.

PE,

as

the

most

flexible

and

promising

new

means

of

precision

genome

editing,

still

suffers

from

low

efficiency,

insufficient

ability

to

manipulate

large

fragments,

complex

design

of

system

components

(such

as

pegRNAs),

incomp