...simpler to implement in genome-wide screens over cDNA libraries. However, using CRISPRa to target individual...highlight a key advantage of CRISPRa over traditional cDNA overexpression: isoform-specific gene expression...— at least, more straightforward than generating cDNA for each gene isoform individually. To highlight...amenable to FACS.  Thus, the researchers inserted human CD8, a gene that encodes a surface protein for which...application can be found here.  Why CRISPRa? Although cDNA overexpression approaches may still be preferable...strategies) and then consider whether traditional cDNA overexpression is suited to your experimental goals...

...large scale, broadly available collection of mutant cDNA clones corresponding to cancer variants found in... genome sequencing studies (as well as reference cDNA clones mapping to well-annotated cellular pathways...Addgene. These include 634 sequence confirmed mutant cDNA clones corresponding to cancer variants derived ...paper Systematic morphological profiling of human gene and allele function via Cell Painting. Rohban...

...they want to activate, we would recommend using a cDNA overexpression vector rather than going through ...genetic screening. gRNAs targeting every gene in the human genome for example, can be made easily and cheaply...protein structure and function (Pan et al., 2018). cDNA libraries, which consist of plasmids that over-express...and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to study in cis regulation and also...

...expression construct, to cells/organisms. This is often cDNA, shRNA, or something similar, and is contained on...infrequent genome integration at the AAVS1 locus in humans. Cap is a structural capsid protein that helps ...-293) during production. Fun fact: There are 57 human adenovirus types, but most vectors are based on ...

...myeloma cell. This RNA was then used to generate cDNA, which  was used as a template for PCR amplification...-mAbs is that it can easily be engineered. Like humans, mice express multiple subclasses of IgG antibody...

...harbor locus” of the human genome, as shown by Dalvai et al., who used HDR to insert cDNA constructs into ... Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. ...

... 3 mCherry 587nm/ 610 nm human β-Actin Actin 4 Citrine 516nm/ 529nm human α-Tubulin Tubulin Repeatable...of these species and can often be used to clone cDNAs from them. These enzymes might not, however, be ...

...desired constructs: lentiviral expression of the human KRAS gene in mammalian cells. STEP 1: Generate an...different ways to generate our desired entry clone - human KRAS flanked by attL sites.  Method A: recombination...types of DNA fragments may be cloned: PCR fragments, cDNA or Genomic DNA and is available for all kind of ...

...BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader... along with other viruses that cause disease in humans such as measles, mumps, and rabies. Research on...

...genome. In humans, if a transposon jumps into a gene, it could be mutagenic and result in human disease....prokaryotes and eukaryotes, and make up ~45% of the human genome (Munoz-Lopez et al., 2010).  So why do transposons.... SB is active in vertebrates and integrates in human cells at rates similar to retroviral vectors. The..., plant, insect, and mammalian cells, including human cells. piggyBac is biased to integrate at transcription...Beauty, piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive...vertebrates and has lower efficiency of integration in human cells than piggyBac and Sleeping Beauty. Minimal...for lentivirus). Large cargos, such as the ~11 kB cDNA for dystrophin, the gene mutated in muscular dystrophy...

...easily isolated from donors. Apart from fibroblasts, human keratinocytes from hair pluck, peripheral blood ...types have been successfully differentiated from human iPSCs. Below is a list of some of these cell types... Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007. 131(...Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007. 318(5858): p. 1917... et al., Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive...PubMed Central PMCID: PMC3161531. 7. Yu, J., et al., Human induced pluripotent stem cells free of vector and...to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell...

...proteins are made by connecting its DNA code to the cDNA encoding for another protein. Effectively, it equips... al. "Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7...

... They contain only a eukaryotic promoter and the cDNA(s) to be expressed and they don’t integrate or replicate... of these functional tests aren’t possible with human iPSCs, the the ability to generate teratomas is ...Robert Lanza, and Kwang-Soo Kim. "Generation of Human Induced Pluripotent Stem Cells by Direct Delivery...and Mahendra S. Rao. "A Review of the Methods for Human iPSC Derivation." Methods in Molecular Biology Pluripotent... Wu. "A nonviral minicircle vector for deriving human iPS cells." Nature Methods 7, no. 3 (2010): 197-...reprogramming factors: Mouse Expression Vectors Human Expression Vectors Carey et al Hockemeyer et al...

...sybody candidates are then reverse transcribed into cDNA fragments that are efficiently subcloned into the...Enzyme Variants. In: Methods in Molecular Biology. Humana Press, pp 133–148. https://doi.org/10.1007/978-...

...constitutive (CMV, SV40, SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30...CreERT2 gene deletion, (3) simultaneous expression of cDNAs and shRNAs with a reporter gene, or (4) multiple...thermophilus which have been used for gene editing in human cells without cross-talk in cognate sgRNA binding...understanding the spatio-temporal regulation of human genes. The plasmids are all available at Addgene...These three enzymes were chosen because they cut human and mouse coding regions infrequently; any MluI,...understanding the role of long noncoding RNAs (lncRNAs) in human health and disease. A primary question in their ...He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison of different...

... both human and fly versions. Arnold, et al. Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral...   Human kinase domain constructs for automated bacterial expression The misregulation of human kinases...Lab recently compiled the Human Kinase Domain Constructs Kit, a library of human kinase domain constructs...gRNAs targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences...repository that can be used to perform screens in human cells. General information regarding how to use ...receptors (GPCRs) yet one third of the GPCRs in the human genome are orphan receptors meaning their ligands...allows researchers to interrogate the druggable human GPCR-ome using a quick, affordable, and ubiquitous...

...grandparents' slow growth period." European Journal of Human Genetics 10.11 (2002): 682-688. PubMed PMID:12404098...reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27 histone methylation...

...Using the ACE system, the Harris lab showed that human APOBEC3A-Cas9n-UGI and APOBEC3B-Cas9n-UGI base editing... is available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). ...

...screening for functional regulatory elements in the human genome. Nature Biotechnology 35:561–568 . https:...mammalian expression constructs including pcDNA-dCas9-p300 Core and pcDNA3.3-Nm-dCas9-p300 Core, as well as pLV-dCas9... in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation and activate...Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a ...