The CRISPR-Cas Mechanism for Adaptive Immunity and Alternate Bacterial Functions Fuels Diverse Biotechnologies DOI Creative Commons
Sydney Newsom,

Hari Priya Parameshwaran,

Lindsie Martin

et al.

Frontiers in Cellular and Infection Microbiology, Journal Year: 2021, Volume and Issue: 10

Published: Jan. 28, 2021

Bacterial and archaeal CRISPR-Cas systems offer adaptive immune protection against foreign mobile genetic elements (MGEs). This function is regulated by sequence specific binding of CRISPR RNA (crRNA) to target DNA/RNA, with an additional requirement a flanking DNA motif called the protospacer adjacent (PAM) in certain systems. In this review, we discuss how same fundamental mechanism RNA-DNA and/or RNA-RNA complementarity utilized bacteria regulate two distinct functions: ward off intruding materials modulate diverse physiological functions. The best documented examples alternate functions are bacterial virulence, biofilm formation, adherence, programmed cell death, quorum sensing. While extensive between crRNA targeted seems constitute efficient phage system, partial be key for several characterized Cas proteins also involved sequence-specific non-specific cleavage control transcriptional regulator expression, mechanisms which still elusive. Over past decade, RNA-guided targeting auxiliary have been transformed into powerful gene editing biotechnological tools. We provide synopsis technologies review. Even abundant mechanistic insights biotechnology tools that currently available, discovery new types holds promise future technological innovations, will pave way precision genome medicine.

Language: Английский

The promise and challenge of therapeutic genome editing DOI
Jennifer A. Doudna

Nature, Journal Year: 2020, Volume and Issue: 578(7794), P. 229 - 236

Published: Feb. 12, 2020

Language: Английский

Citations

907
CRISPR/Cas Systems towards Next-Generation Biosensing DOI
Yi Li, Shiyuan Li, Jin Wang

et al.

Trends in biotechnology, Journal Year: 2019, Volume and Issue: 37(7), P. 730 - 743

Published: Jan. 15, 2019

Language: Английский

Citations

781
Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing DOI Creative Commons
James K. Nuñez, Jin Chen, Greg C. Pommier

et al.

Cell, Journal Year: 2021, Volume and Issue: 184(9), P. 2503 - 2519.e17

Published: April 1, 2021

Language: Английский

Citations

519
Genome-Scale Imaging of the 3D Organization and Transcriptional Activity of Chromatin DOI Creative Commons
Jun-Han Su, Pu Zheng, Seon Kinrot

et al.

Cell, Journal Year: 2020, Volume and Issue: 182(6), P. 1641 - 1659.e26

Published: Aug. 20, 2020

Language: Английский

Citations

476
CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future DOI Creative Commons

Fathema Uddin,

Charles M. Rudin, Triparna Sen

et al.

Frontiers in Oncology, Journal Year: 2020, Volume and Issue: 10

Published: Aug. 7, 2020

A series of recent discoveries harnessing the adaptive immune system prokaryotes to perform targeted genome editing is having a transformative influence across biological sciences. The discovery Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) proteins has expanded applications genetic research in thousands labs globe redefining our approach gene therapy. Traditional therapy raised some concerns, as its reliance on viral vector delivery therapeutic transgenes can cause both insertional oncogenesis immunogenic toxicity. By obliviating concerns by traditional therapy, CRISPR technology provides relatively simple efficient alternative for site-specific editing. Although it apparent advantages, CRISPR/Cas9 brings own set limitations which must be addressed safe clinical translation. This review focuses evolution role shifting paradigm. We emerging data trials consider best strategy move forward with this powerful but still new technology.

Language: Английский

Citations

408
High-content CRISPR screening DOI Open Access
Christoph Bock, Paul Datlinger, Florence M. Chardon

et al.

Nature Reviews Methods Primers, Journal Year: 2022, Volume and Issue: 2(1)

Published: Feb. 10, 2022

Language: Английский

Citations

381
Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing DOI Creative Commons
Xiaoshu Xu, Augustine Chemparathy, Leiping Zeng

et al.

Molecular Cell, Journal Year: 2021, Volume and Issue: 81(20), P. 4333 - 4345.e4

Published: Sept. 3, 2021

Compact and versatile CRISPR-Cas systems will enable genome engineering applications through high-efficiency delivery in a wide variety of contexts. Here, we create an efficient miniature Cas system (CasMINI) engineered from the type V-F Cas12f (Cas14) by guide RNA protein engineering, which is less than half size currently used CRISPR (Cas9 or Cas12a). We demonstrate that CasMINI can drive high levels gene activation (up to thousands-fold increases), while natural fails function mammalian cells. show has comparable activities Cas12a for activation, highly specific, allows robust base editing editing. expect be broadly useful cell therapy ex vivo vivo.

Language: Английский

Citations

286
The CRISPR-Cas toolbox and gene editing technologies DOI Creative Commons

Guanwen Liu,

Qiupeng Lin, Shuai Jin

et al.

Molecular Cell, Journal Year: 2021, Volume and Issue: 82(2), P. 333 - 347

Published: Dec. 29, 2021

Language: Английский

Citations

264
High-content CRISPR screening DOI
Christoph Bock, Paul Datlinger, Florence M. Chardon

et al.

Nature Reviews Methods Primers, Journal Year: 2022, Volume and Issue: 2(1)

Published: Feb. 10, 2022

Language: Английский

Citations

209
Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs delivered by adeno-associated virus DOI Creative Commons

Do Yon Kim,

Jeong Mi Lee,

Su Bin Moon

et al.

Nature Biotechnology, Journal Year: 2021, Volume and Issue: 40(1), P. 94 - 102

Published: Sept. 2, 2021

Abstract Gene therapy would benefit from a miniature CRISPR system that fits into the small adeno-associated virus (AAV) genome and has high cleavage activity specificity in eukaryotic cells. One of most compact CRISPR-associated nucleases yet discovered is archaeal Un1Cas12f1. However, Un1Cas12f1 its variants have very low In present study, we redesigned natural guide RNA at five sites: 5′ terminus trans -activating (tracrRNA), tracrRNA–crRNA complementary region, penta(uridinylate) sequence, 3′ crRNA disordered stem 2 region tracrRNA. These optimizations synergistically increased average indel frequency by 867-fold. The optimized enabled efficient, specific editing human cells when delivered plasmid vectors, PCR amplicons AAV. As cleaves outside protospacer, it can be used to create large deletions efficiently. engineered showed efficiency comparable SpCas9 similar AsCas12a.

Language: Английский

Citations

201