Full-Spectrum Targeted Mutagenesis in Plant and Animal Cells
Abstract
:1. Introduction
2. Directed Mutagenesis Using CRISPR Nucleases
3. Diversifying Base Editors
4. EvolvR
5. Other Approaches for Targeted Mutagenesis
6. In Situ and Ex Vivo Directed Evolution Using Targeted Mutagenesis
7. Limitations and Future Prospects
8. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AAG | Alkyl adenine DNA glycosylase |
ABE | Adenine base editor |
ACC | Acetyl-coenzyme A carboxylase |
AID | Activation-induced cytidine deaminase |
AIDmono | Monomeric AID |
AP site | Apurinic/apyrimidinic site |
APOBEC3A | Apolipoprotein B mRNA editing enzyme catalytic polypeptide 3A |
APOBEC3B | Apolipoprotein B mRNA editing enzyme catalytic polypeptide 3B |
BE | Base editor |
BER | Base excision repair |
BFP | Blue fluorescent protein |
Cas | CRISPR-associated protein |
CBE | Cytosine base editor |
CGBE | C-to-G base editor |
CRISPR | Clustered regularly interspaced short palindromic repeats |
dCas9 | Catalytically dead Cas9 |
DSB | Double-strand break |
GESTALT | Genome editing of synthetic target arrays for lineage tracing |
GFP | Green fluorescent protein |
gRNA | Guide RNA |
HDR | Homology-directed repair |
Indels | Insertions and deletions |
MEK1 kinase | Mitogen-activated protein kinase kinase 1 |
MEMOIR | Memory by engineered mutagenesis with optical in situ readout |
MPG | N-methylpurine glycosylase |
mSCRIBE | Mammalian synthetic cellular recorders integrating biological events |
MSH2 | MutS homolog 2 |
nCas9 | Cas9 nickase |
NHEJ | Non-homologous end joining |
ORF | Open reading frame |
PAM | Protospacer adjacent motif |
PmCDA1 | Petromyzon marinus cytidine deaminase 1 |
rAPOBEC1 | Rat apolipoprotein B mRNA editing enzyme catalytic polypeptide 1 |
scRNA-seq | Single-cell RNA sequencing |
sgRNA | Single guide RNA |
SPACE | Synchronous programmable adenine and cytosine editor |
ssDNA | Single-stranded DNA |
STEME | Saturated targeted endogenous mutagenesis editor |
TAM | Targeted AID-mediated mutagenesis |
TLS | Translesion synthesis |
TRACE | T7 polymerase-driven continuous editing |
UGI | Uracil glycosylase inhibitor |
UNG | Uracil-DNA glycosylase |
XRCC1 | X-ray repair cross-complementing protein 1 |
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Deaminase | Hotspot 1 | Editing Profile | Editing Profile (With UGI or in UGN/MSH2 Double Mutant) | Reference |
---|---|---|---|---|
hAIDx (P182X) | WGC | C/G to all nucleotides, with a preference for C/G to T/A | C/G to T/A | [15,23] |
hAID*Δ | WRC | [15] | ||
AIDmono | WRC, AGCT | |||
PmCDA1 | WRC | |||
APOBEC3A (A3A) | TCA, TC, TCC (using nCas9) | |||
APOBEC3B mutant (A3BAct) | TCA, TC | |||
rAPOBEC1 | TC | |||
AID-3C (AIDmono with APOBEC3C substrate-recognition loop) | TTC | |||
AID-3F (AIDmono with APOBEC3F substrate-recognition loop) | TGC |
Target Protein | Evolution Goal | Evolution System | Biological System | Identified Major Mutations | Reference |
---|---|---|---|---|---|
Anti-hapten 4-hydroxy-3-nitrophenyl acetyl antibody B1-8 | Improve antigen affinity | CRISPR-X with AIDmono/APOBEC3A | HEK293T cells | W33L, T30I, T30S, S31R, T58I, A97G | [15] |
Human monoclonal antibody A2Ab | Improve antigen affinity | CRISPR-X with AID*Δ | HEK 293 cells | D74H, W102 L, M112I, G121D, R124P | [34] |
30S ribosomal proteins S5 and S12 | Spectinomycin and streptomycin resistance | EvolvR | E. coli | Δ17–19, K23N and Δ24, Δ24, Δ26, G27D (rpsE) | [31] |
BCR-ABL | Imatinib (Gleevec) resistance | TAM | Human K562 cells | T315I, T319I | [23] |
Mitogen-activated protein kinase kinase 1 (MEK1 kinase) | Selumetinib and trametinib resistance | TRACE | Human A375 cells | E38K, V211D | [32] |
Acetyl-coenzyme A carboxylase (ACC) | Herbicide (haloxyfop) resistance | STEMES | Rice | W2125C, P1927F, S1866F, A1884P | [35] |
Blue fluorescent protein (BFP) | Shift fluorescence spectrum to that of green fluorescent protein (GFP) | TRACE | HEK293T cells | H66Y | [32] |
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Iaffaldano, B.; Reiser, J. Full-Spectrum Targeted Mutagenesis in Plant and Animal Cells. Int. J. Mol. Sci. 2021, 22, 857. https://doi.org/10.3390/ijms22020857
Iaffaldano B, Reiser J. Full-Spectrum Targeted Mutagenesis in Plant and Animal Cells. International Journal of Molecular Sciences. 2021; 22(2):857. https://doi.org/10.3390/ijms22020857
Chicago/Turabian StyleIaffaldano, Brian, and Jakob Reiser. 2021. "Full-Spectrum Targeted Mutagenesis in Plant and Animal Cells" International Journal of Molecular Sciences 22, no. 2: 857. https://doi.org/10.3390/ijms22020857