ArticlePloS one2014
Simple and efficient methods for enrichment and isolation of endonuclease modified cells.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 36 citations in OpenAlex.
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- A Gene Transfer-Positive Cell Sorting System Utilizing Membrane-Anchoring Affinity Tag.Frontiers in bioengineering and biotechnology · 2022Article
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- Optimization of AsCas12a for combinatorial genetic screens in human cells.Nature biotechnology · 2021Article
- Loss of MAR1 Function is a Marker for Co-Selection of CRISPR-Induced Mutations in Plants.Frontiers in genome editing · 2021Article
- Strategies for High-Efficiency Mutation Using the CRISPR/Cas System.Frontiers in cell and developmental biology · 2021Review
- An efficient and precise method for generating knockout cell lines based on CRISPR-Cas9 system.Engineering in life sciences · 2020Article
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- EditR: A Method to Quantify Base Editing from Sanger Sequencing.The CRISPR journal · 2018Article
- MicroRNA-Based Attenuation of Influenza Virus across Susceptible Hosts.Journal of virology · 2018Article
- Comparative analysis of chimeric ZFP-, TALE- and Cas9-piggyBac transposases for integration into a single locus in human cells.Nucleic acids research · 2017Article
- Prosurvival long noncoding RNAeLife · 2017Article
- Marker-free coselection for CRISPR-driven genome editing in human cells.Nature methods · 2017Article
- Efficient Precision Genome Editing in iPSCs via Genetic Co-targeting with Selection.Stem cell reports · 2017Article
Corrections and comments
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Authors and funding
11 authors at 3 institutions in 1 country.
Funding
Abstract
The advent of Transcription Activator-Like Effector Nucleases (TALENs), and similar technologies such as CRISPR, provide a straightforward and cost effective option for targeted gene knockout (KO). Yet, there is still a need for methods that allow for enrichment and isolation of modified cells for genetic studies and therapeutics based on gene modified human cells. We have developed and validated two methods for simple enrichment and isolation of single or multiplex gene KO's in transformed, immortalized, and human progenitor cells. These methods rely on selection of a phenotypic change such as resistance to a particular drug or ability to grow in a selective environment. The first method, termed co-transposition, utilizes integration of a piggyBac transposon vector encoding a drug resistance gene. The second method, termed co-targeting, utilizes TALENs to KO any gene that when lost induces a selectable phenotype. Using these methods we also show removal of entire genes and demonstrate that TALENs function in human CD34+ progenitor cells. Further, co-transposition can be used to generate conditional KO cell lines utilizing an inducible cDNA rescue transposon vector. These methods allow for robust enrichment and isolation of KO cells in a rapid and efficient manner.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.