Use of Lentiviral Vectors to Deliver and Express Bicistronic Transgenes in Developing Chicken Embryos
Open accessAbstract
The abilities of lentiviral vectors to carry large transgenes (∼8kb) and to efficiently infect and integrate these genes into the genomes of both dividing and non-dividing cells make them ideal candidates for transport of genetic material into cells and tissues. Given the properties of these vectors, it is somewhat surprising that they have seen only limited use in studies of developing tissues and in particular of the developing nervous system. Over the past several years, we have taken advantage of the large capacity of these vectors to explore the expression characteristics of several dual promoter and 2A peptide bicistronic transgenes in developing chick neural retina, with the goal of identifying transgene designs that reliably express multiple proteins in infected cells. Here we summarize the activities of several of these transgenes in neural retina and provide detailed methodologies for packaging lentivirus and delivering the virus into the developing neural tubes of chicken embryos in ovo, procedures that have been optimized over the course of several years of use in our laboratory. Conditions to hatch injected embryos are also discussed. The chicken-specific techniques will be of highest interest to investigators using avian embryos, development and packaging of lentiviral vectors that reliably express multiple proteins in infected cells should be of interest to all investigators whose experiments demand manipulation and expression of multiple proteins in developing cells and tissues.
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Lentiviruses can carry genes of up to 8 kb.
“There are other viral vectors that have a larger packaging capacity, such as Lentiviruses (LVs), which can carry genes of up to 8 kb [11]”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [11]
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- Authors
- Semple-Rowland, S. L., & Berry, J.
- Affiliations
- University of Florida (US)
- Venue
- Methods
- Publisher
- Elsevier BV
- Published
- 2014-04
- Volume
- 66
- Issue
- 3
- Pages
- 466-473
- ISSN
- 1046-2023 (ISSN-L)
- PMC
- PMC3823667
- OpenAlex
- W2073030837
- Cited by
- 23
- References
- 59
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 1 · 2022: 1 · 2021: 3 · 2019: 1 · 2018: 1 · 2017: 8 · 2016: 1 · 2015: 1 · 2014: 3
- FWCI
- 1.10
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- NIH (EY11388)
- NIH Vision Core (EY08571)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Keywords
- lentiviral vectors, 2A peptide, dual promoter, chick embryo, neural retina, neural tube, in ovo injection, transgene expression, avian embryo, embryonic development
- MeSH terms
- Animals, Chick Embryo, Genetic Engineering, Genetic Vectors, Retina, Lentivirus, Transgenes, Animals, Genetically Modified, Embryonic Development
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