Recent advances in CRISPR-based genome editing technology and its applications in cardiovascular research DOI Creative Commons
Zhenhua Li, Jun Wang, Jingping Xu

et al.

Military Medical Research, Journal Year: 2023, Volume and Issue: 10(1)

Published: March 10, 2023

Abstract The rapid development of genome editing technology has brought major breakthroughs in the fields life science and medicine. In recent years, clustered regularly interspaced short palindromic repeats (CRISPR)-based toolbox been greatly expanded, not only with emerging CRISPR-associated protein (Cas) nucleases, but also novel applications through combination diverse effectors. Recently, transposon-associated programmable RNA-guided systems have uncovered, adding myriads potential new tools to toolbox. CRISPR-based revolutionized cardiovascular research. Here we first summarize advances involving newly identified Cas orthologs, engineered variants systems, then discuss CRISPR-Cas precise editing, such as base prime editing. We highlight progress research using technologies, including generation genetically modified vitro animal models diseases (CVD) well treating different types CVD. Finally, current limitations future prospects technologies are discussed.

Language: Английский

CRISPR-based genome editing through the lens of DNA repair DOI Creative Commons

Tarun S. Nambiar,

Lou Baudrier,

Pierre Billon

et al.

Molecular Cell, Journal Year: 2022, Volume and Issue: 82(2), P. 348 - 388

Published: Jan. 1, 2022

Language: Английский

Citations

154
Structural biology of CRISPR–Cas immunity and genome editing enzymes DOI
Joy Y. Wang, Patrick Pausch, Jennifer A. Doudna

et al.

Nature Reviews Microbiology, Journal Year: 2022, Volume and Issue: 20(11), P. 641 - 656

Published: May 13, 2022

Language: Английский

Citations

135
Glycerol Additive Boosts 100-fold Sensitivity Enhancement for One-Pot RPA-CRISPR/Cas12a Assay DOI
Lin Mei,

Huahua Yue,

Tian Tian

et al.

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(23), P. 8277 - 8284

Published: May 30, 2022

CRISPR/Cas12, a highly efficient and specific nucleic acid recognition system, has been broadly employed to detect amplified DNA products. However, most reported methods adopt two-step detection mode that needs liquid transfer step, thus complicating the procedure posing risk of aerosol contamination. A one-pot method can obviate these problems, but it suffers from poor efficiency due loss amplification templates elicited by CRISPR/Cas12 cleavage. In this study, we discovered glycerol additive dramatically promoted recombinase polymerase (RPA)-CRISPR/Cas12a method. Compared with glycerol-free version, its sensitivity was nearly 100-fold higher close canonical Further investigation displayed enhanced attributed phase separation RPA CRISPR/Cas12a system during initial reaction caused viscosity. This triumphantly harnessed for African swine fever virus (ASFV) SARS-CoV-2, achieving naked-eye readout through smartphone-equipped device. The currently developed glycerol-enhanced RPA-CRISPR/Cas12a be an advantageous point-of-care platform on account simplicity, high sensitivity, universality.

Language: Английский

Citations

123
Genetics of adaptation DOI Creative Commons
Kirsten Bomblies, Catherine L. Peichel

Proceedings of the National Academy of Sciences, Journal Year: 2022, Volume and Issue: 119(30)

Published: July 18, 2022

The rediscovery of Mendel's work showing that the heredity phenotypes is controlled by discrete genes was followed reconciliation Mendelian genetics with evolution natural selection in middle last century Modern Synthesis. In past two decades, dramatic advances genomic methods have facilitated identification loci, genes, and even individual mutations underlie phenotypic variants are putative targets selection. Moreover, these also changed how we can study adaptation flipping problem around, allowing us to first examine what loci show evidence having been under selection, then connecting genetic variation. As a result, now an expanding list actual changes potentially adaptive Here, synthesize considering effects context cellular environments, genomes, organisms, populations has provided new insights architecture adaptation.

Language: Английский

Citations

75
Drug delivery systems for CRISPR-based genome editors DOI
Victoria J. Madigan, Feng Zhang, James E. Dahlman

et al.

Nature Reviews Drug Discovery, Journal Year: 2023, Volume and Issue: 22(11), P. 875 - 894

Published: Sept. 18, 2023

Language: Английский

Citations

74
Light‐Start CRISPR‐Cas12a Reaction with Caged crRNA Enables Rapid and Sensitive Nucleic Acid Detection DOI

Menglu Hu,

Ruhan Liu, Zhiqiang Qiu

et al.

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 62(23)

Published: April 5, 2023

Abstract The clustered regularly interspaced short palindromic repeats (CRISPR) system is a promising platform for nucleic acid detection. Regulating the CRISPR reaction would be extremely useful to improve detection efficiency and speed of diagnostic applications. Here, we have developed light‐start CRISPR‐Cas12a by employing caged RNA (crRNA). When combined with recombinase polymerase amplification, robust photocontrolled one‐pot assay achieved. simpler 50‐fold more sensitive than conventional assay. This improved also facilitates development faster method. clinical samples demonstrated that 10–20 min sufficient effective detection, which much current gold‐standard technique PCR. We expect this advance in diagnostics promote its widespread applications biomedicine, agriculture, food safety.

Language: Английский

Citations

70
Adenine transversion editors enable precise, efficient A•T-to-C•G base editing in mammalian cells and embryos DOI
Liang Chen,

Mengjia Hong,

Changming Luan

et al.

Nature Biotechnology, Journal Year: 2023, Volume and Issue: 42(4), P. 638 - 650

Published: June 15, 2023

Language: Английский

Citations

69
Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes DOI Creative Commons
Qian Liu,

Chenglu Zhao,

Kai Sun

et al.

Molecular Plant, Journal Year: 2023, Volume and Issue: 16(3), P. 616 - 631

Published: Feb. 7, 2023

Language: Английский

Citations

61
Targeted genome-modification tools and their advanced applications in crop breeding DOI
Boshu Li, Chao Sun, Jiayang Li

et al.

Nature Reviews Genetics, Journal Year: 2024, Volume and Issue: 25(9), P. 603 - 622

Published: April 24, 2024

Language: Английский

Citations

55
Prediction of prime editing insertion efficiencies using sequence features and DNA repair determinants DOI Creative Commons
Jonas Koeppel, Juliane Weller, Elin Madli Peets

et al.

Nature Biotechnology, Journal Year: 2023, Volume and Issue: 41(10), P. 1446 - 1456

Published: Feb. 16, 2023

Most short sequences can be precisely written into a selected genomic target using prime editing; however, it remains unclear what factors govern insertion. We design library of 3,604 various lengths and measure the frequency their insertion four sites in three human cell lines, different editor systems varying DNA repair contexts. find that length, nucleotide composition secondary structure sequence all affect rates. also discover 3' flap nucleases TREX1 TREX2 suppress longer sequences. Combining features machine learning model, we predict relative insertions site with R = 0.70. Finally, demonstrate how our accurate prediction user-friendly software help choose codon variants common fusion tags insert at high efficiency, provide catalog empirically determined rates for over hundred useful

Language: Английский

Citations

49