The discovery and development of Cas12A
DOI:
https://doi.org/10.54097/beta8y02Keywords:
High Specificity; T-rich PAM sequence; Off-target effects; Multi-target Editing; crRNA array.Abstract
CRISPR-Cas12a stands as a transformative tool in genome editing, offering unparalleled precision and efficiency. Its utilization of an RNA-guided Cas12a enzyme enables targeted double-strand DNA breaks, allowing for the manipulation of genes with remarkable accuracy. Notable features such as high specificity, multi-target editing capabilities, and a variety of Cas12a variants provide researchers with a versatile toolkit for diverse applications. From aiding in drug discovery through large-scale genetic screens to enabling precise corrections of disease-causing mutations in gene therapy, CRISPR-Cas12a holds immense potential across various fields. However, challenges such as off-target effects, lower editing efficiency compared to Cas9, and financial limitations pose significant considerations. Stringent biosecurity measures and ethical dilemmas surrounding germline editing and accessibility further underscore the need for cautious advancement. Nonetheless, CRISPR-Cas12a continues to pave the way for transformative advancements in biotechnology, medicine, and our understanding of biology.
Downloads
References
I. Grissa, G. Vergnaud, C. Pourcel. Clustered regularly interspaced short palindromic repeats (CRISPRs) for the genotyping of bacterial pathogens. Methods Mol Biol, 2009, 551:105-116.
S.A. Shah, S. Erdmann, F.J. Mojica, R.A. Garrett. Protospacer recognition motifs: mixed identities and functional diversity. RNA Biol, 2013, 10(5):891-899.
B. Paul, G. Montoya. CRISPR-Cas12a: Functional overview and applications. Biomed J, 2020, 43(1):8-17.
F.B. Ayanoğlu, A.E. Elçin, Y.M. Elçin. Bioethical issues in genome editing by CRISPR-Cas9 technology. Turk J Biol 2020, 44(2):110-120.
A. Regalado. CRISPR inventor Feng Zhang calls for moratorium on gene-edited babies, MIT Technology Review, 2018, November 26.
L. Marks. Professor Jennifer Doudna, What is Biotechnology? Oct 2017.
S. Begley. Watch: Meet one of the world’s most groundbreaking scientists. He’s 34. STAT News, Nov. 6, 2015.
S. Khan, E. Sallard. Current and Prospective Applications of CRISPR-Cas12a in Pluricellular Organisms. Mol Biotechnol, 2023, 65(2):196-205.
H. Kim, W.J. Lee, Y. Oh, S.H. Kang, J.K. Hur, H. Lee, W. Song, K.S. Lim, Y.H. Park, B.S. Song, Y.B. Jin, B.H. Jun, C. Jung, D.S. Lee, S.U. Kim, S.H. Lee. Enhancement of target specificity of CRISPR-Cas12a by using a chimeric DNA-RNA guide. Nucleic Acids Res, 2020, 48(15):8601-8616.
J.P. Magnusson, A.R. Rios, L. Wu, L.S. Qi. Enhanced Cas12a multi-gene regulation using a CRISPR array separator. Elife, 2021, 10.
X. Ling, L. Chang, H. Chen, X. Gao, J. Yin, Y. Zuo, Y. Huang, B. Zhang, J. Hu, T. Liu. Improving the efficiency of CRISPR-Cas12a-based genome editing with site-specific covalent Cas12a-crRNA conjugates. Molecular Cell, 2021, 81(22):4747-4756.e4747.
K. Lee, Y. Zhang, B.P. Kleinstiver, J.A. Guo, M.J. Aryee, J. Miller, A. Malzahn, S. Zarecor, C.J. Lawrence-Dill, J.K. Joung, Y. Qi, K. Wang. Activities and specificities of CRISPR/Cas9 and Cas12a nucleases for targeted mutagenesis in maize. Plant Biotechnol J, 2019, 17(2):362-372.
R.A. Gier, K.A. Budinich, N.H. Evitt, Z. Cao, E.S. Freilich, Q. Chen, J. Qi, Y. Lan, R.M. Kohli, J. Shi. High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening. Nature Communications, 2020, 11(1):3455.
N. Pagliarulo. Pricey new gene therapies for sickle cell pose access test, BioPharma Dive, Dec. 8, 2023.
Y. Chen, X. Ma, L. Pan, S. Yang, X. Chen, F. Wang, D. Yang, M. Li, P. Wang. A CRISPR-Cas12a-based assay for one-step preamplification-free detection of viral DNA. Sensors and Actuators B: Chemical, 2024, 399:134813.
P. Schindele, H. Puchta. Engineering CRISPR/LbCas12a for highly efficient, temperature-tolerant plant gene editing. Plant Biotechnol J, 2020, 18(5):1118-1120.
M.A. Collingwood, C.A. Vakulskas, L. Zhang, R. Turk, N.M. Bode, A.M. Jacobi, M.S. McNeill, K. McQuisten, B. Thommandru, M.A. Behlke. Engineered AsCas12a Variants with Enhanced Activity and Broadened PAM Compatibility. J Biomol Tech, 2019, 30(Suppl):S49.
X. Liu, X. Liu, C. Zhou, J. Lv, X. He, Y. Liu, H. Xie, B. Wang, X. Lv, L. Tang, M. Li, C. Liu, J. Zhao, Y. Liu, Z. Song, F. Gu. Engineered FnCas12a with enhanced activity through directional evolution in human cells. J Biol Chem, 2021, 296:100394.
K.E. Watters, C. Fellmann, H.B. Bai, S.M. Ren, J.A. Doudna. Systematic discovery of natural CRISPR-Cas12a inhibitors. Science, 2018, 362(6411):236-239.
E. Ma, K. Chen, H. Shi, E.C. Stahl, B. Adler, M. Trinidad, J. Liu, K. Zhou, J. Ye, J.A. Doudna. Improved genome editing by an engineered CRISPR-Cas12a. Nucleic Acids Res, 2022, 50(22):12689-12701.
G. Rubeis, F. Steger. Risks and benefits of human germline genome editing: An ethical analysis. Asian Bioeth Rev, 2018, 10(2):133-141.
MLA style: Emmanuelle Charpentier – Facts – 2020, Nobel Prize Outreach AB 2024, 4 Apr 2024.
Eugenics and Scientific Racism, National Human Genome Research Institute, 2022, May 18.
Downloads
Published
Issue
Section
License
Copyright (c) 2024 Highlights in Science, Engineering and Technology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
