Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives

Overview

Journal Viruses

Publisher MDPI

Specialty Microbiology

Date 2021 Aug 10

PMID 34372494

Citations 42

Authors

Wendy Dong

Boris Kantor

Affiliations

Soon will be listed here.

Abstract

CRISPR/Cas technology has revolutionized the fields of the genome- and epigenome-editing by supplying unparalleled control over genomic sequences and expression. Lentiviral vector (LV) systems are one of the main delivery vehicles for the CRISPR/Cas systems due to () its ability to carry bulky and complex transgenes and () sustain robust and long-term expression in a broad range of dividing and non-dividing cells in vitro and in vivo. It is thus reasonable that substantial effort has been allocated towards the development of the improved and optimized LV systems for effective and accurate gene-to-cell transfer of CRISPR/Cas tools. The main effort on that end has been put towards the improvement and optimization of the vector's expression, development of integrase-deficient lentiviral vector (IDLV), aiming to minimize the risk of oncogenicity, toxicity, and pathogenicity, and enhancing manufacturing protocols for clinical applications required large-scale production. In this review, we will devote attention to () the basic biology of lentiviruses, and () recent advances in the development of safer and more efficient CRISPR/Cas vector systems towards their use in preclinical and clinical applications. In addition, we will discuss in detail the recent progress in the repurposing of CRISPR/Cas systems related to base-editing and prime-editing applications.

Citing Articles

Chimeric Antigen Receptor Cell Therapy: Empowering Treatment Strategies for Solid Tumors.

Jaing T, Hsiao Y, Wang Y Curr Issues Mol Biol. 2025; 47(2).

PMID: 39996811 PMC: 11854309. DOI: 10.3390/cimb47020090.

RNA Structure: Past, Future, and Gene Therapy Applications.

Haseltine W, Hazel K, Patarca R Int J Mol Sci. 2025; 26(1.

PMID: 39795966 PMC: 11719923. DOI: 10.3390/ijms26010110.

Advances in CRISPR-Cas technology and its applications: revolutionising precision medicine.

Azeez S, Hamad R, Hamad B, Shekha M, Bergsten P Front Genome Ed. 2024; 6:1509924.

PMID: 39726634 PMC: 11669675. DOI: 10.3389/fgeed.2024.1509924.

Safer and efficient base editing and prime editing via ribonucleoproteins delivered through optimized lipid-nanoparticle formulations.

Holubowicz R, Du S, Felgner J, Smidak R, Choi E, Palczewska G Nat Biomed Eng. 2024; 9(1):57-78.

PMID: 39609561 PMC: 11754100. DOI: 10.1038/s41551-024-01296-2.

CRISPR/Cas9 Ribonucleoprotein Nucleofection for Genome Editing in Primary Human Keratinocytes: Knockouts, Deletions, and Homology-Directed Repair Mutagenesis.

Bamundo M, Palumbo S, DAuria L, Missero C, Di Girolamo D Curr Protoc. 2024; 4(11):e70056.

PMID: 39601181 PMC: 11600394. DOI: 10.1002/cpz1.70056.

References

Kantor B, Bayer M, Ma H, Samulski J, Li C, McCown T . Notable reduction in illegitimate integration mediated by a PPT-deleted, nonintegrating lentiviral vector. Mol Ther. 2010; 19(3):547-56. PMC: 3048187. DOI: 10.1038/mt.2010.277. View

Koblan L, Doman J, Wilson C, Levy J, Tay T, Newby G . Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction. Nat Biotechnol. 2018; 36(9):843-846. PMC: 6126947. DOI: 10.1038/nbt.4172. View

Ortinski P, ODonovan B, Dong X, Kantor B . Integrase-Deficient Lentiviral Vector as an All-in-One Platform for Highly Efficient CRISPR/Cas9-Mediated Gene Editing. Mol Ther Methods Clin Dev. 2017; 5:153-164. PMC: 5424571. DOI: 10.1016/j.omtm.2017.04.002. View

Iwakuma T, Cui Y, Chang L . Self-inactivating lentiviral vectors with U3 and U5 modifications. Virology. 1999; 261(1):120-32. DOI: 10.1006/viro.1999.9850. View

Lewis P, Emerman M . Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus. J Virol. 1994; 68(1):510-6. PMC: 236313. DOI: 10.1128/JVI.68.1.510-516.1994. View

Craigie R, Fujiwara T, Bushman F . The IN protein of Moloney murine leukemia virus processes the viral DNA ends and accomplishes their integration in vitro. Cell. 1990; 62(4):829-37. DOI: 10.1016/0092-8674(90)90126-y. View

Chen V, Moncalvo M, Tringali D, Tagliafierro L, Shriskanda A, Ilich E . The mechanistic role of alpha-synuclein in the nucleus: impaired nuclear function caused by familial Parkinson's disease SNCA mutations. Hum Mol Genet. 2020; 29(18):3107-3121. PMC: 7645704. DOI: 10.1093/hmg/ddaa183. View

Naldini L, Blomer U, Gage F, Trono D, Verma I . Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci U S A. 1996; 93(21):11382-8. PMC: 38066. DOI: 10.1073/pnas.93.21.11382. View

Pickar-Oliver A, Gersbach C . The next generation of CRISPR-Cas technologies and applications. Nat Rev Mol Cell Biol. 2019; 20(8):490-507. PMC: 7079207. DOI: 10.1038/s41580-019-0131-5. View

10.

Pattanayak V, Lin S, Guilinger J, Ma E, Doudna J, Liu D . High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nat Biotechnol. 2013; 31(9):839-43. PMC: 3782611. DOI: 10.1038/nbt.2673. View

11.

Zennou V, Petit C, Guetard D, Nerhbass U, Montagnier L, Charneau P . HIV-1 genome nuclear import is mediated by a central DNA flap. Cell. 2000; 101(2):173-85. DOI: 10.1016/S0092-8674(00)80828-4. View

12.

Chavez A, Scheiman J, Vora S, Pruitt B, Tuttle M, Iyer E . Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015; 12(4):326-8. PMC: 4393883. DOI: 10.1038/nmeth.3312. View

13.

Choi J, Dang Y, Abraham S, Ma H, Zhang J, Guo H . Lentivirus pre-packed with Cas9 protein for safer gene editing. Gene Ther. 2016; 23(7):627-33. DOI: 10.1038/gt.2016.27. View

14.

Leavitt A, Rose R, Varmus H . Both substrate and target oligonucleotide sequences affect in vitro integration mediated by human immunodeficiency virus type 1 integrase protein produced in Saccharomyces cerevisiae. J Virol. 1992; 66(4):2359-68. PMC: 289031. DOI: 10.1128/JVI.66.4.2359-2368.1992. View

15.

Ihry R, Worringer K, Salick M, Frias E, Ho D, Theriault K . p53 inhibits CRISPR-Cas9 engineering in human pluripotent stem cells. Nat Med. 2018; 24(7):939-946. DOI: 10.1038/s41591-018-0050-6. View

16.

Konermann S, Brigham M, Trevino A, Joung J, Abudayyeh O, Barcena C . Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2014; 517(7536):583-8. PMC: 4420636. DOI: 10.1038/nature14136. View

17.

Cockrell A, Ma H, Fu K, McCown T, Kafri T . A trans-lentiviral packaging cell line for high-titer conditional self-inactivating HIV-1 vectors. Mol Ther. 2006; 14(2):276-84. DOI: 10.1016/j.ymthe.2005.12.015. View

18.

Komor A, Zhao K, Packer M, Gaudelli N, Waterbury A, Koblan L . Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity. Sci Adv. 2017; 3(8):eaao4774. PMC: 5576876. DOI: 10.1126/sciadv.aao4774. View

19.

Shalem O, Sanjana N, Hartenian E, Shi X, Scott D, Mikkelson T . Genome-scale CRISPR-Cas9 knockout screening in human cells. Science. 2013; 343(6166):84-87. PMC: 4089965. DOI: 10.1126/science.1247005. View

20.

Komor A, Badran A, Liu D . Editing the Genome Without Double-Stranded DNA Breaks. ACS Chem Biol. 2017; 13(2):383-388. PMC: 5891729. DOI: 10.1021/acschembio.7b00710. View